Journal: Cancers

Article Title: Novel Function of Cancer Stem Cell Marker ALDH1A3 in Glioblastoma: Pro-Angiogenesis through Paracrine PAI-1 and IL-8.

doi: 10.3390/cancers15174422

Figure Lengend Snippet: Figure 1. Upregulation of ALDH1A3 in oxGBM cells is associated with the increased expression and release of pro-angiogenic factors PAI-1 and IL-8. GBM cell lines were transduced with ALDH1A3 for overexpression (oxGBM) or with empty vector (evGBM). (A) Confirmation of up-regulation of ALDH1A3 mRNA level in oxGBM cells by RT2-PCR. (B) Confirmation of up-regulation of ALDH1A3 protein expression by Western blot. wt, wild-type cells. The uncropped blots are shown in Figure S7; (C) Angiogenesis array. The blots showed duplicated dots for 55 angiogenesis-related proteins in the media of oxU373 or evU373. 10 of 55 proteins were upregulated more than 2-fold in ox group compared to ev group (indicated by rectangle). They are: (1) Ang-1, (2) artemin, (3) TF, (4) ET-1, (5) GM-CSF, (6) IL-8, (7) PDGF-AA, (8) PAI-1, (9) PEDF, and (10) uPA. (D) Semi-quantification of the dots representing PAI-1 and IL-8. (E) Immunofluorescence staining of GBM cells. U373 (left panel) and LN229 (right panel). Co-localization of ALDH1A3 with PAI-1 and IL-8 was observed in oxGBM cells, whereas no immunoreactivity of PAI-1 and IL-8 was detected in evGBM cells. (F) mRNA expression of PAI-1 and IL-8 in transduced U373 cells and the effect of inhibitors. Tiplaxtinin (Tip, 30 µM) and reparixin (Rep, 1 µM) are the specific inhibitors of PAI-1 and IL-8 receptors CXCR1/2, respectively. (G) Detection of PAI-1 and potential signaling proteins by Western blot. IOD: optical density. The uncropped blots are shown in Figure S8. **, p < 0.01; ***, p < 0.001, compared with ev. ##, p < 0.01; ###, p < 0.001, compared with ox.

Article Snippet: To evaluate the potential angiogenic factors released from oxGBMs, an array was carried out using a human angiogenesis array kit (cat# ARY007; R&D Systems, Wiesbaden, Germany) according to the manufacturer’s protocol.

Techniques: Expressing, Transduction, Over Expression, Plasmid Preparation, Western Blot, Staining

Journal: Cancers

Article Title: Novel Function of Cancer Stem Cell Marker ALDH1A3 in Glioblastoma: Pro-Angiogenesis through Paracrine PAI-1 and IL-8.

doi: 10.3390/cancers15174422

Figure Lengend Snippet: Figure 2. Overexpression of ALDH1A3 in GBM cells activated endothelial angiogenesis in indirect co-culture with endothelial cells (ECs), which was reversed by treatment with respective inhibitors of PAI-1 or IL-8 receptors CXCR1/2. Indirect co-culture was performed by culture of HBMECs in a conditioned medium (CM) containing the media derived from evGBMs or oxGBMs and ECGM in a ratio of 1:1. PAI-1 inhibitor tiplaxtinin (Tip, 30 µM) and CXCR1/2 inhibitor reparixin (Rep, 1 µM) or vehicle DMSO (0.1%) was added to CM, followed by EC behavior study. All data were reproduced in three independent experiments. (A) Proliferation assay in HBMEC and HUVEC. Indirect co-culture of HBMECs and HUVECs with CM derived from oxU373 and oxLN229 stimulated EC proliferation, which was completely reversed by the treatment of tiplaxtinin, not by reparixin. (B) Scratch assay in HBMEC. Left panel: images were acquired 24 h after scratching. Scale bar: 200 µm. Right panel: quantitative analysis. Culture of HBMECs with CM derived from oxU373 or oxLN229 (oxCM) significantly promoted HBMEC migration, which was reversed by the treatment of tiplaxtinin and reparixin, respectively. (C) Transwell invasion assay in HBMEC. Left panel: Representative images of invaded cells were acquired after 24 h of incubation. Scale bar: 100 µm. Right panel: quantitative analysis. Culture of HBMECs with oxCM accelerated HBMEC invasion. This effect was significantly inhibited by the treatment of reparixin but not by tiplaxtinin. (D) Tube formation assay in HBMEC. Left panel: representative images of tube formation. Scale bar: 200 µm. Right panel: quantitative analysis of branching points per field. Tube formation in HBMECs was stimulated by incubation with oxCM, which was completely diminished by both inhibitors. (E) Sprouting assay in HBMEC. Left panel: representative images of sprouting in HBMECs after 24 h of co-culture. Scale bar: 100 µm. A pronounced increase in sprouting was observed in HBMECs cultured in oxCM. Tiplaxtinin and reparixin suppressed the sprouting effect resulting from oxCM. *, p < 0.05; **, p < 0.01 and ***, p < 0.001, compared with evCM. #, p < 0.05; ##, p < 0.01 and ###, p < 0.001, compared with oxCM.

Article Snippet: To evaluate the potential angiogenic factors released from oxGBMs, an array was carried out using a human angiogenesis array kit (cat# ARY007; R&D Systems, Wiesbaden, Germany) according to the manufacturer’s protocol.

Techniques: Over Expression, Co-Culture Assay, Derivative Assay, Proliferation Assay, Wound Healing Assay, Migration, Transwell Invasion Assay, Incubation, Tube Formation Assay, Cell Culture

Journal: Cancers

Article Title: Novel Function of Cancer Stem Cell Marker ALDH1A3 in Glioblastoma: Pro-Angiogenesis through Paracrine PAI-1 and IL-8.

doi: 10.3390/cancers15174422

Figure Lengend Snippet: Figure 4. oxGBM-derived culture media stimulated angiogenesis on CAM, which was fully rescued by the treatment of tiplaxtinin and reparixin. CAM was incubated with the culture media derived from evU373 or oxU373 cells with or without tiplaxtinin (Tip, 30 µM) or reparixin (Rep, 1 µM) or DMSO (as a vehicle control, 0.1%) for 72 h. (A) Microscopy view of the vasculature structure on CAM. More enriched microvessel network was clearly visible in the ox group, which was significantly reduced in tiplaxtinin- and reparixin-treated CAMs. The images were acquired using a stereo microscope on ED13 (scale bar: 1 mm). Stem vessel (arrow); branched microvessel network (arrow heads). (B) Histological features of CAM after hematoxylin-eosin (H&E) staining. The CAM consists of the chorionic epithelium layer (ChE), allantoic epithelium (AE) layer, and the mesenchymal (MES) layer (arrows). Microvessel (arrowheads) density was much higher in the MES layer of ox section compared to ev section, which was clearly reduced in tiplaxtinin- and reparixin-treated sections. Scale bar: 50 µm. (C) Quantitative analysis of branching point of vessels based on microscopy images. The number of branching points and microvessels was counted by the ImageJ software (v1.1.53t) in 3 fields/CAM (n = 10 CAM/group). (D) Quantitative analysis of microvessel numbers based on H&E-stained CAM sections. Microvessel number was counted manually on H&E-stained CAMs. 10 fields/section (n = 6 sections/group). ***, p < 0.001, compared with ev. ###, p < 0.001, compared with ox.

Article Snippet: To evaluate the potential angiogenic factors released from oxGBMs, an array was carried out using a human angiogenesis array kit (cat# ARY007; R&D Systems, Wiesbaden, Germany) according to the manufacturer’s protocol.

Techniques: Derivative Assay, Incubation, Control, Microscopy, Staining, Software

Journal: Cancer gene therapy

Article Title: Mc-hES, a novel plasmid carrying human endostatin gene, inhibits nasopharyngeal carcinoma growth.

doi: 10.1038/cgt.2011.72

Figure Lengend Snippet: Figure 2 Expression profiles and biological function of endostatin in eukaryotic cells. (a) RT–PCR analysis of endostatin transcripts in CNE-2 cells transfected with plasmid. Lane 1, DL2000, bands of 2, 1, 0.75, 0.5 and 0.25 kb; lane 2, mc-hES; lane 3, pcDNA-hES; lane 4, pj-hES; lane 5, p2jc31; lane 6, empty control; lane 7, positive control. (b) Western blot assay. Lane 1, p2jc31; lane 2, mc-hES; lane 3, pcDNA-hES; lane 4, pj-hES. (c) Expression profiles of endostatin in the supernatant of cells transfected with plasmids. Columns represent the mean of three independent experiments, each conducted in triplicate; bars, s.e. Po 0.05, compared with pj-hES and pcDNA-hES treated group. (d) MTT assay. HUVECs were treated with CM from transfected CNE-2 cells and cell proliferation was evaluated after 72 h. Columns, cell proliferation normalized against p2jc31 control (mean±s.d.). Mc-hES versus pcDNA-hES, P ¼ 0.019; mc-hES versus pj-hES, P ¼ 0.005; pcDNA-hES versus p2jc31, P ¼ 0.237. (e) Tube formation assay. HUVECs were treated with CM. After 16 h of incubation images of tube formation were captured and tube formation was scored in one 50 microscopic field. The number of tubes formed was quantitated and the data are presented as the mean±s.d. per field ( 50). Data were normalized to p2jc31-treated control (n ¼ 5). Mc-hES versus pcDNA-hES, P ¼ 0.015; mc-hES versus pj-hES, P ¼ 0.008; pcDNA-hES versus p2jc31, P ¼ 0.009. (f) Migration assay. HUVECs were treated with CM and pipetted into inserts of matrigel-coated transwells. Addition of chemoattractant, HUVECs in CM containing 50 ng ml1 vascular endothelial growth factor, to the lower well of boyden chamber stimulated endothelial migration to the underside of the transwell membrane. After 6 h of incubation, migrated cells were stained by DAPI. The number of cells that migrated was counted by microscopy and the data are presented as the mean±s.d. per field ( 100). Data were normalized to p2jc31-treated control (n ¼ 5). Mc-hES versus pcDNA-hES, P ¼ 0.01; mc-hES versus pj-hES, P ¼ 0.015; pcDNA-hES versus p2jc31, P ¼ 0.019.

Article Snippet: CNE-2 cell supernatants were harvested as conditioned medium (CM) after 72 h of transfection and endostatin expression was evaluated using a human endostatin ELISA kit (R&D systems, Minneapolis, MN) according to the recommendation of the manufacturer.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection, Plasmid Preparation, Control, Positive Control, Western Blot, MTT Assay, Tube Formation Assay, Incubation, Migration, Membrane, Staining, Microscopy

Journal: Cancer gene therapy

Article Title: Mc-hES, a novel plasmid carrying human endostatin gene, inhibits nasopharyngeal carcinoma growth.

doi: 10.1038/cgt.2011.72

Figure Lengend Snippet: Figure 4 (a) Endostatin expression profile in the tumor tissues. For expression profiles of endostatin, results are given in nanograms per mg of tumor tissue after single administration. Columns, mean of three mice; bars, s.d. mc-hES versus pcDNA-hES. Po0.001 at days 3, 7, 14 and 20. (b) Results from representative experiments are shown for RT–PCR analysis of endostatin transcripts. b-actin was used as loading control. b, blank; M, DL2000; , negative control; þ , positive control; s, saline.

Article Snippet: CNE-2 cell supernatants were harvested as conditioned medium (CM) after 72 h of transfection and endostatin expression was evaluated using a human endostatin ELISA kit (R&D systems, Minneapolis, MN) according to the recommendation of the manufacturer.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Negative Control, Positive Control, Saline

Journal: Thoracic Cancer

Article Title: Knockdown of circ‐RAD23B inhibits non‐small cell lung cancer progression via the miR ‐142‐3p/ MAP4K3 axis

doi: 10.1111/1759-7714.14319

Figure Lengend Snippet: Circ‐RAD23B knockdown blocked NSCLC development in vitro. (a) The expression of circ‐RAD23B in NSCLC tissues and normal tissues was measured by qPCR. (b) The expression of circ‐RAD23B in H1299, A549 and 16 HBE cells was detected by qPCR. (c) The efficiency of circ‐RAD23B interference was checked by qPCR. (d) The effect of circ‐RAD23B knockdown on cell viability was examined by CCK‐8 assay. (e) The effect of circ‐RAD23B knockdown on cell proliferation was examined by colony formation assay. (f and g) The effect of circ‐RAD23B knockdown on migration and invasion was examined by transwell assay. (h) Matrigel tube formation assay was performed to evaluate the angiogenic ability of HUVECs. (i) The effect of circ‐RAD23B knockdown on cell cycle distribution was determined by flow cytometry assay. (j) The effect of circ‐RAD23B knockdown on cell apoptosis was analyzed by flow cytometry assay. (k) Expression of PCNA and Bax affected by circ‐RAD23B knockdown was detected by western blot. * p < 0.05

Article Snippet: An Annexin V‐FITC apoptosis detection kit (Beyotime) was utilized to monitor cell apoptosis.

Techniques: Knockdown, In Vitro, Expressing, CCK-8 Assay, Colony Assay, Migration, Transwell Assay, Tube Formation Assay, Flow Cytometry, Western Blot

Journal: Thoracic Cancer

Article Title: Knockdown of circ‐RAD23B inhibits non‐small cell lung cancer progression via the miR ‐142‐3p/ MAP4K3 axis

doi: 10.1111/1759-7714.14319

Figure Lengend Snippet: Circ‐RAD23B knockdown blocked NSCLC development in vitro by mediating miR‐142‐3p. In H1299 and A549 cells transfected with si‐circ‐RAD23B‐1 or si‐circ‐RAD23B‐1 + miR‐142‐3p inhibitor, (a and b) cell proliferation was assessed by CCK‐8 assay and colony formation assay. (c and d) Cell migration and cell invasion were assessed by transwell assay. (e) The angiogenic ability was assessed by Matrigel tube formation assay. (f and g) Cell cycle distribution was determined by flow cytometry assay. (h) Cell apoptosis was analyzed by flow cytometry assay. (i and j) The expression of PCNA and Bax was measured by western blot. * p < 0.05

Article Snippet: An Annexin V‐FITC apoptosis detection kit (Beyotime) was utilized to monitor cell apoptosis.

Techniques: Knockdown, In Vitro, Transfection, CCK-8 Assay, Colony Assay, Migration, Transwell Assay, Tube Formation Assay, Flow Cytometry, Expressing, Western Blot

Journal: Thoracic Cancer

Article Title: Knockdown of circ‐RAD23B inhibits non‐small cell lung cancer progression via the miR ‐142‐3p/ MAP4K3 axis

doi: 10.1111/1759-7714.14319

Figure Lengend Snippet: MiR‐142‐3p blocked NSCLC development in vitro by depleting MAP4K3. In H1299 and A549 cells transfected with miR‐142‐3p mimic or miR‐142‐3p mimic+pc‐MAP4K3, (a and b) cell proliferation was assessed by CCK‐8 assay and colony formation assay. (c and d) cell migration and cell invasion were assessed by transwell assay. (e) The angiogenic ability was investigated by Matrigel tube formation assay. (f and g) Cell cycle distribution was determined by flow cytometry assay. (h) Cell apoptosis was investigated by flow cytometry assay. (i and j) The expression of PCNA and Bax was detected by western blot. * p < 0.05

Article Snippet: An Annexin V‐FITC apoptosis detection kit (Beyotime) was utilized to monitor cell apoptosis.

Techniques: In Vitro, Transfection, CCK-8 Assay, Colony Assay, Migration, Transwell Assay, Tube Formation Assay, Flow Cytometry, Expressing, Western Blot

Journal: The Kaohsiung journal of medical sciences

Article Title: Paeoniflorin inhibits colorectal cancer cell stemness through the miR-3194-5p/catenin beta-interacting protein 1 axis.

doi: 10.1002/kjm2.12736

Figure Lengend Snippet: FIGURE 1 PF suppressed CRC cell viability, migration, invasion, stemness, and EMT by deactivating Wnt/β-catenin signaling. HCT116 and SW480 cells were treated with PF (0, 5, 10, 30, and 60 μM). (A) Cell viability was measured using CCK-8 assay. (B–C) Cell migration and invasion were measured using transwell assays. (D–E) β-Catenin, p-β-catenin, Axin2, C-myc, OCT4, Nanog, and Sox2 levels were analyzed using western blot assay. (F) Sphere-formation capacity was measured using the sphere-formation assay. (G) Protein levels of MMP2, MMP9, E-cadherin, and vimentin in CRC cells were determined using western blot assay. The measurement data were presented as mean ± SD. All data were obtained from at least three replicate experiments. *p < 0.05, **p < 0.01, ***p < 0.001. Axin2, axis inhibition protein 2; CCK-8, cell counting kit-8; CRC, colorectal cancer; EMT, epithelial–mesenchymal transition; MMP2, matrix metalloproteinase-2; MMP9, metalloproteinase-9; PF, paeoniflorin; OCT4, octamer binding factor 4; Sox2, SRY-box transcription factor 2.

Article Snippet: Normal human colon epithelial cells and human CRC cells (HCT116 and SW480) were obtained from ATCC (VA, USA) and cultured in DMEM (Gibco, CA, USA) containing 10% fetal bovine serum (Gibco) at 37 C with 5% CO2.

Techniques: Migration, CCK-8 Assay, Western Blot, Tube Formation Assay, Inhibition, Cell Counting, Binding Assay

Journal: The Kaohsiung journal of medical sciences

Article Title: Paeoniflorin inhibits colorectal cancer cell stemness through the miR-3194-5p/catenin beta-interacting protein 1 axis.

doi: 10.1002/kjm2.12736

Figure Lengend Snippet: FIGURE 2 MiR-3194-5p enhanced CRC cell viability, migration, invasion, EMT, and stemness. (A) qRT-PCR was employed to detect miR- 3194-5p expression in CRC cells. We induced miR-3194-5p knockdown/overexpression in CRC cells by transfecting miR-3194-5p inhibitor/ mimics into the cells. (B) miR-3194-5p expression in cells was examined using qRT-PCR. (C) CCK-8 assay was conducted to examine cell viability. (D–E) Cell migration and invasion were measured using transwell assays. (F) Protein levels of MMP2, MMP9, E-cadherin, and vimentin in CRC cells were measured using western blot assay. (G) OCT4, Nanog, and Sox2 protein levels in CRC cells were measured using western blot assay. (H) Sphere-formation capacity was determined using the sphere-formation assay. The measurement data were presented as mean ± SD. All data were obtained from at least three replicate experiments. *p < 0.05, **p < 0.01, ***p < 0.001. CCK-8, cell counting kit-8; CRC, colorectal cancer; EMT, epithelial–mesenchymal transition; MMP2, matrix metalloproteinase-2; MMP9, metalloproteinase-9; OCT4, octamer binding factor 4; qRT- PCR, quantitative real-time polymerase chain reaction; Sox2, SRY-box transcription factor 2.

Article Snippet: Normal human colon epithelial cells and human CRC cells (HCT116 and SW480) were obtained from ATCC (VA, USA) and cultured in DMEM (Gibco, CA, USA) containing 10% fetal bovine serum (Gibco) at 37 C with 5% CO2.

Techniques: Migration, Quantitative RT-PCR, Expressing, Knockdown, Over Expression, CCK-8 Assay, Western Blot, Tube Formation Assay, Cell Counting, Binding Assay, Real-time Polymerase Chain Reaction

Journal: Experimental Hematology & Oncology

Article Title: Ubiquitin ligase subunit FBXO9 inhibits V-ATPase assembly and impedes lung cancer metastasis

doi: 10.1186/s40164-024-00497-4

Figure Lengend Snippet: FBXO9-mediated ATP6V1A ubiquitination impairs metastasis of lung cancer cells. A , B Ubiquitination assays were performed in HEK293T cells by co-transfection with full-length HA-ATP6V1A, HA-ATP6V1A-KR (lysine residues removed), HA-ATP6V1A-K393 (K393 reintroduced into the ATP6V1A-KR mutant), or HA-ATP6V1A-K393R (single lysine substitution mutant), in the presence or absence of FLAG-FBXO9. After 48 h, cells were lysed, followed by Ni-NTA bead pull-down and immunoblotting for HA-ATP6V1A. C Alignment of ATP6V1A K393 conservation across species. D Ubiquitination analysis of K393 in HA-Atp6v1a by co-expressing Flag-Fbxo9 and His-Ub with indicated HA-Atp6v1a constructs (WT, K393, or K393R) in HEK293T cells. Cells were lysed after 48 h, followed by Ni-NTA bead pull-down and immunoblotting for HA-Atp6v1a. E V-ATPase complex reconstitution was achieved in H1299 cells by introducing sgRNA-resistant ATP6V1A (wild-type or K393R mutant) using a Tet-Off system. CRISPR was then used to deplete endogenous ATP6V1A, allowing the V-ATPase complex to be reconstituted. Immunoblot analysis confirmed comparable expression levels of the added ATP6V1A and endogenous subunits ATP6V1B and ATP6V1D1, in the experimental group compared to the control cells, after 48 h of doxycycline treatment. F–I Recombinant cell lines H1299-KO-V1A-wt, H1299-KO-V1A-K393R, and H1299-EGFP-sgCtrl were used in a transwell migration assay (F, G) and tumor sphere formation assay (H, I) to evaluate the effects of ATP6V1A ubiquitination on cell migration and tumor sphere growth( P < 0.001). J–N To assess the effects of ATP6V1A ubiquitination on cell migration and tumor sphere growth, V1A-23aa was introduced into A549 cells to inhibit ATP6V1A ubiquitination (J) which was followed by transwell migration assay (K, L) and tumor sphere formation assay (M, N) to evaluate the impact on cell migration and tumor sphere growth, respectively ( P < 0.001). O, P Impact of ATP6V1A ubiquitination inhibition by FBXO9 on in vivo lung metastasis was assessed. A549 cells expressing V1A-23 aa were injected into mice via the tail vein. After 6 weeks, the lungs of the mice were harvested for examination and H&E staining. Metastasis nodules were quantified and statistically analyzed using fluorescence microscopy. Error bars represent the mean ± SD. **** P < 0.0001 indicates the significance level

Article Snippet: Subsequently, the puromycin-resistant cells were treated with doxycycline for 72 h. Furthermore, for the knockdown of human FBXO9, cancer cells were transiently transfected with siRNA oligonucleotides using Lipofectamine RNAiMAX (Thermo Fisher), following the manufacturer’s guidelines.

Techniques: Ubiquitin Proteomics, Cotransfection, Mutagenesis, Western Blot, Expressing, Construct, CRISPR, Control, Recombinant, Transwell Migration Assay, Tube Formation Assay, Migration, Inhibition, In Vivo, Injection, Staining, Fluorescence, Microscopy

Journal: Frontiers in pharmacology

Article Title: Homocysteine Impairs Endothelial Cell Barrier Function and Angiogenic Potential via the Progranulin/EphA2 Pathway.

doi: 10.3389/fphar.2020.614760

Figure Lengend Snippet: FIGURE 1 | Comparison and correlation analysis of clinical indexes between the CCS patients (n 36) and the ACS patients (n 37). (A–D) Comparison of serum levels of EphA2, PGRN, VCAM1 and Hcy, respectively. CCS: chronic coronary syndrome; ACS: acute coronary syndrome. EphA2: Eph-receptor tyrosine kinase-type A2, PGRN: Progranulin, VCAM1: vascular cell adhesion molecule-1; Hcy: homocysteine.

Article Snippet: The cells were stimulated with DL-homocysteine (Sigma Aldrich) 2.0 mM in the presence or absence of recombinant human progranulin (rhPGRN) protein 200 ng/mL (R&D systems) for 24 h as well as treated with a variety of concentrations of rhPGRN (0, 50, 100, 200, 400 ng/mL) for 24 h depending on the experimental goals.

Techniques: Comparison

Journal: Frontiers in pharmacology

Article Title: Homocysteine Impairs Endothelial Cell Barrier Function and Angiogenic Potential via the Progranulin/EphA2 Pathway.

doi: 10.3389/fphar.2020.614760

Figure Lengend Snippet: FIGURE 5 | RhPGRN ameliorated the Hcy-induced reduction in cell viability and migration. (A) Results of cell viability in HUVECs after stimulated with 2.0 mM Hcy in the presence or absence of 200 ng/mL rhPGRN (B,C) Results of the scratch wound assay were recorded and calculated at 12 and 24 h after stimulated with 2.0 mM Hcy in the presence or absence of 200 ng/mL rhPGRN. Scale bar, 200 μm. (D) The tube formation assay of HUVECs stimulated with 2.0 mM Hcy, 200 ng/mL rhPGRN, and Hcy + rhPGRN for 6 h. Scale bar, 200 μm (E,F) The cell migration was evaluated by the Transwell cell migration assay in HUVECs stimulated with 2.0 mM Hcy, 200 ng/mL rhPGRN, and Hcy + rhPGRN for 6∼8 h. Scale bar, 100 μm. (G) Western blotting analysis of the protein levels of VCAM1 and VE-cadherin stimulated with 2.0 mM Hcy in the presence or absence of 200 ng/mL rhPGRN for 24 h. Data are presented as means ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001 vs. vehicle control, ANOVA, n 3 or 6).

Article Snippet: The cells were stimulated with DL-homocysteine (Sigma Aldrich) 2.0 mM in the presence or absence of recombinant human progranulin (rhPGRN) protein 200 ng/mL (R&D systems) for 24 h as well as treated with a variety of concentrations of rhPGRN (0, 50, 100, 200, 400 ng/mL) for 24 h depending on the experimental goals.

Techniques: Migration, Scratch Wound Assay Assay, Tube Formation Assay, Cell Migration Assay, Western Blot, Control

Journal: Frontiers in pharmacology

Article Title: Homocysteine Impairs Endothelial Cell Barrier Function and Angiogenic Potential via the Progranulin/EphA2 Pathway.

doi: 10.3389/fphar.2020.614760

Figure Lengend Snippet: FIGURE 6 | Effects of rhPGRN combined with shRNA interference of EphA2 on adhesion molecules and cytokines and NF-κB p65 and AKT pathways in Hcy- injured HUVECs. (A) Cells interfered with EphA2-shRNA (EphA2-RNAi) or NC-shRNA (NC-RNAi), and then the EphA2-RNAi group was treated with Hcy 2.0 mM, 200 ng/mL rhPGRN, Hcy + rhPGRN for 24 h. Representative Western blotting results of protein levels of VE-cadherin and VCAM1 between groups. (B) The summarized data of the protein levels of VCAM1 and VE-cadherin (C) Representative Western blotting gel documents and summarized data showing the levels of phospho-AKT (Ser473) and total AKT. (D) Representative western blot gel documents and summarized data showing the levels of p-NF-κB p65 (Ser536) and total NF-κB p65. Data are presented as means ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001 vs. NC-RNAi group, ANOVA, n 3). NC: negative control.

Article Snippet: The cells were stimulated with DL-homocysteine (Sigma Aldrich) 2.0 mM in the presence or absence of recombinant human progranulin (rhPGRN) protein 200 ng/mL (R&D systems) for 24 h as well as treated with a variety of concentrations of rhPGRN (0, 50, 100, 200, 400 ng/mL) for 24 h depending on the experimental goals.

Techniques: shRNA, Western Blot, Negative Control

Journal: Frontiers in pharmacology

Article Title: Homocysteine Impairs Endothelial Cell Barrier Function and Angiogenic Potential via the Progranulin/EphA2 Pathway.

doi: 10.3389/fphar.2020.614760

Figure Lengend Snippet: FIGURE 7 | The effect of exogenous rhPGRN on the PGRN/EphA2 axis and the NF-κB p65 and AKT pathways. (A) HUVECs were administered with a variety of concentrations of rhPGRN (0, 50, 100, 200, 400 ng/mL) for 24 h. Western blotting gel documents of p-EphA2 (Ser897), EphA2, and PGRN showed relative protein levels in different groups. (B) The summarized data of the protein levels of EphA2 and PGRN (C) The summarized data of the protein levels of fold p-EphA2 (Ser897). (D) Phospho-AKT (Ser473), total AKT, NF-κB p65 (Ser536), and total NF-κB p65 signaling molecules were detected by the Western blotting. (E) The summarized data the levels phospho-NF-κB p65 (Ser536), total NF-κB p65. Data are presented as means ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001 vs. vehicle, ANOVA, n 3).

Article Snippet: The cells were stimulated with DL-homocysteine (Sigma Aldrich) 2.0 mM in the presence or absence of recombinant human progranulin (rhPGRN) protein 200 ng/mL (R&D systems) for 24 h as well as treated with a variety of concentrations of rhPGRN (0, 50, 100, 200, 400 ng/mL) for 24 h depending on the experimental goals.

Techniques: Western Blot

Journal: Molecular Endocrinology

Article Title: Targeting Androgen Receptor to Suppress Macrophage-induced EMT and Benign Prostatic Hyperplasia (BPH) Development

doi: 10.1210/me.2012-1079

Figure Lengend Snippet: The co-culture with macrophages can increase prostate epithelial cell growth in a 3D culture model. BPH-1 cells were cultured in a 3D culture condition with the co-cultured medium of BPH-1/THP-1 cells or control medium. A (magnification, ×10), The co-cultured medium of BPH-1/THP-1 cells increased the sphere diameter of BPH-1 cells (scale bar, 20 μm), as shown in gross observation of BPH-1 spheres under mono-culture (upper) and co-culture (lower). B, Results are expressed as the average size of the spheres (sphere number: 30 for each experiment set) for three independent experiments. The diameter difference of spheres was analyzed by two-tailed Student's t test. C, (magnification, ×100), mPrE cells were cultured in a 3D culture condition, and the results of the sphere diameter of mPrE cells were expressed as described in panel B; scale bar, 100 μm.

Article Snippet: Cell culture and co-culture experiments Human benign prostate epithelial cell line, BPH-1, was purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Co-Culture Assay, Cell Culture, Control, Two Tailed Test

Journal: Molecular Endocrinology

Article Title: Targeting Androgen Receptor to Suppress Macrophage-induced EMT and Benign Prostatic Hyperplasia (BPH) Development

doi: 10.1210/me.2012-1079

Figure Lengend Snippet: The coculture with macrophages induces EMT gene expression in prostate epithelial cells in 3D culture condition. A, Quantitative PCR analysis of EMT markers in BPH-1 cells. N-cadherin, Snail, and TGF-β2 were increased after co-culture. E-Cadherin expression was decreased. Bars represent an average expression level (fold) of three independent experiments. Error bars represent sem. *, P < 0.01. The difference of gene expression was analyzed by two-tailed Student's t test. B, Western blot analysis of EMT markers in BPH-1 cells. C, Western blot analysis of EMT markers, Snail and N-cadherin, in mPrE cells with or without coculture with RAW264.7 macrophages.

Article Snippet: Cell culture and co-culture experiments Human benign prostate epithelial cell line, BPH-1, was purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Co-Culture Assay, Expressing, Two Tailed Test, Western Blot

Journal: Molecular Endocrinology

Article Title: Targeting Androgen Receptor to Suppress Macrophage-induced EMT and Benign Prostatic Hyperplasia (BPH) Development

doi: 10.1210/me.2012-1079

Figure Lengend Snippet: The expression of AR in prostate epithelial cells increased THP-1 cell migration, sphere number, and EMT induction. A, The expression of AR in BPH cells enhanced the migration of THP-1 cells during co-culture. The THP-1 migration assay was performed as described in Fig. 2. Briefly, THP-1 cells were co-cultured with BPH-V or BPH-AR cells. Bars represent an average number of migrated cells of three independent experiments (left panel; **, P = 0.003). The sphere formation assay was performed as described in Fig. 3 to examine the 3D growth of BPH-V and BPH-AR cells (middle panel; *, P = 0.0022). Results are shown as described in Fig. 3. AR expression was confirmed by Western blot analysis (right panel). B, Quantitative PCR and Western blot analysis of Snail, N-cadherin, and E-cadherin expression in BPH-v and BPH-AR cells (left and middle panels); *, P < 0.01. Western blot analysis of EMT markers in mPrE-v and mPrE-AR cells after coculture (right panel). C, ASC-J9 can suppress AR expression in BPH-AR cells and inhibit BPH-AR sphere formation in the presence of coculture medium. P values are shown as indicated. D, ASC-J9 can block induction of EMT markers in BPH-AR cells. DMSO, Dimethylsulfoxide.

Article Snippet: Cell culture and co-culture experiments Human benign prostate epithelial cell line, BPH-1, was purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Expressing, Migration, Co-Culture Assay, Cell Culture, Tube Formation Assay, Western Blot, Real-time Polymerase Chain Reaction, Blocking Assay

Journal: Oncology reports

Article Title: Knockdown of Bmi1 inhibits the stemness properties and tumorigenicity of human bladder cancer stem cell-like side population cells.

doi: 10.3892/or.2013.2919

Figure Lengend Snippet: Figure 1. Identification of side population (SP) cells from the human bladder cancer T24 cells. (A) SP cells were sorted by flow cytometry using Hoechst 33342 (Hoe) from T24 cells with or without pretreatment with verapamil (V) for 30 min. (B) Relative mRNA expression of ABCG2 and OCT4 detected in SP and non-SP (NSP) T24 cells by real-time RT-PCR. GAPDH was used as an internal reference. The data are shown as means ± standard deviation (SD). *P<0.05 vs. NSP cells.

Article Snippet: The human bladder cancer cell line T24 [American Type Culture Collection (ATCC), Manassas, VA, USA] was cultured in RMPI-1640 medium (Gibco, Invitrogen, Carlsbad, CA, USA), supplemented with 10% fetal bovine serum (FBS) (Gibco), 100 U/ml penicillin G and 100 μg/ml streptomycin.

Techniques: Flow Cytometry, Expressing, Quantitative RT-PCR, Standard Deviation

Journal: Oncology reports

Article Title: Knockdown of Bmi1 inhibits the stemness properties and tumorigenicity of human bladder cancer stem cell-like side population cells.

doi: 10.3892/or.2013.2919

Figure Lengend Snippet: Figure 2. Bmi1 is highly expressed in SP T24 cells. (A) Relative expression of Bmi1 as detected in SP and NSP cells by real-time RT-PCR. GAPDH was used as internal reference. *P<0.05 vs. NSP cells. (B) Western blot analyses of Bmi1 protein expression in SP and NSP cells. (C) Immunocytochemical analyses of Bmi1 protein expression. The nucleus was stained with DAPI (blue), and Bmi1 was stained with CY3 (red) (magnification, x200). The data are shown as means ± SD.

Article Snippet: The human bladder cancer cell line T24 [American Type Culture Collection (ATCC), Manassas, VA, USA] was cultured in RMPI-1640 medium (Gibco, Invitrogen, Carlsbad, CA, USA), supplemented with 10% fetal bovine serum (FBS) (Gibco), 100 U/ml penicillin G and 100 μg/ml streptomycin.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Staining

Journal: Oncology reports

Article Title: Knockdown of Bmi1 inhibits the stemness properties and tumorigenicity of human bladder cancer stem cell-like side population cells.

doi: 10.3892/or.2013.2919

Figure Lengend Snippet: Figure 3. Effects of Bmi1 knockdown on SP T24 cell phenotype, cell proliferation and migration in vitro. SP cells were sorted by flow cytometry using Hoechst 33342 in T24 cells (A) and purified T24 SP cells (B) after Bmi1 knockdown and long-term culture (10 days). (C) SP and NSP cell proliferation was monitored by MTT assays for 7 days. (D) SP cell proliferation after knockdown of Bmi1, as assessed by the MTT assays for 5 days. *P<0.05, **P<0.01 vs. SP-scramble-siRNA. (E) The cell migration capacity was analyzed by Transwell assays. Five random fields (magnification, x200) were photographed and cells were counted. The data are shown as means ± SD. *P<0.05, **P<0.01.

Article Snippet: The human bladder cancer cell line T24 [American Type Culture Collection (ATCC), Manassas, VA, USA] was cultured in RMPI-1640 medium (Gibco, Invitrogen, Carlsbad, CA, USA), supplemented with 10% fetal bovine serum (FBS) (Gibco), 100 U/ml penicillin G and 100 μg/ml streptomycin.

Techniques: Knockdown, Migration, In Vitro, Flow Cytometry, Purification

Journal: Oncology reports

Article Title: Knockdown of Bmi1 inhibits the stemness properties and tumorigenicity of human bladder cancer stem cell-like side population cells.

doi: 10.3892/or.2013.2919

Figure Lengend Snippet: Figure 4. Bmi1 regulates the self-renewal of SP T24 cells in vitro. (A) Tumor sphere formation assay was used to assess the self-renewal capacity of SP and NSP cells. Scale bar, 100 µm. (B) Self-renewal capacity of SP cells after Bmi1 knockdown. Five random fields (x100) were photographed, and the sphere numbers were counted after 7 days. The data are shown as means ± SD. **P<0.01.

Article Snippet: The human bladder cancer cell line T24 [American Type Culture Collection (ATCC), Manassas, VA, USA] was cultured in RMPI-1640 medium (Gibco, Invitrogen, Carlsbad, CA, USA), supplemented with 10% fetal bovine serum (FBS) (Gibco), 100 U/ml penicillin G and 100 μg/ml streptomycin.

Techniques: In Vitro, Tube Formation Assay, Knockdown

Journal: Oncology reports

Article Title: Knockdown of Bmi1 inhibits the stemness properties and tumorigenicity of human bladder cancer stem cell-like side population cells.

doi: 10.3892/or.2013.2919

Figure Lengend Snippet: Figure 5. Bmi1 knockdown enhances SP T24 cell sensitivity to cisplatin. SP, NSP, SP-Bmi-siRNA and SP-scramble-siRNA cells were treated with various concentrations of cisplatin (0.5, 2 and 4 µg/ml) for 48 h, and the inhibition rate (IR) was detected by MTT assays. Survival rate (SR) (%) = (mean absor bance of the test well/mean absorbance of the control) x 100%. Inhibition rate (IR) was calculated as IR (%) = 100% - SR (%). The data are shown as means ± SD. **P<0.01.

Article Snippet: The human bladder cancer cell line T24 [American Type Culture Collection (ATCC), Manassas, VA, USA] was cultured in RMPI-1640 medium (Gibco, Invitrogen, Carlsbad, CA, USA), supplemented with 10% fetal bovine serum (FBS) (Gibco), 100 U/ml penicillin G and 100 μg/ml streptomycin.

Techniques: Knockdown, Inhibition, Control

Journal: Oncology reports

Article Title: Knockdown of Bmi1 inhibits the stemness properties and tumorigenicity of human bladder cancer stem cell-like side population cells.

doi: 10.3892/or.2013.2919

Figure Lengend Snippet: Figure 6. Bmi1 knockdown inhibits cell cycle progression through derepression of the p16INK4a/p14ARF locus in SP T24 cells. (A) Bmi1, p16INK4a and p14ARF mRNA expression was determined by real-time RT-PCR. GAPDH was used as an internal reference. (B) Bmi1, p16INK4a and p14ARF protein expression was determined by western blotting. β-actin was used as an internal reference. (C) Cell cycle progression in SP cells after Bmi1 knockdown was determined by flow cytometric analysis using propidium iodide (PI) staining. The data are shown as means ± SD. *P<0.05, **P<0.01.

Article Snippet: The human bladder cancer cell line T24 [American Type Culture Collection (ATCC), Manassas, VA, USA] was cultured in RMPI-1640 medium (Gibco, Invitrogen, Carlsbad, CA, USA), supplemented with 10% fetal bovine serum (FBS) (Gibco), 100 U/ml penicillin G and 100 μg/ml streptomycin.

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Western Blot, Staining

Journal: Oncology reports

Article Title: Knockdown of Bmi1 inhibits the stemness properties and tumorigenicity of human bladder cancer stem cell-like side population cells.

doi: 10.3892/or.2013.2919

Figure Lengend Snippet: Figure 7. Bmi1 silencing suppresses the tumorigenicity of SP T24 cells in vivo. (A) SP-Bmi1-siRNA and SP-scramble-siRNA T24 cells (1x105) were subcutane ously injected into the left and right sides of the back of recipient mice, respectively. The mice were sacrificed 6 weeks later. (B) Sizes of the tumors formed from subcutaneously injected SP-scramble-siRNA T24 cells. (C) Results of hematoxylin and eosin (H&E) staining showed that the tumor formed from subcutaneously injected SP-scramble-siRNA T24 cells was a typical urothelium carcinoma (magnification, x100).

Article Snippet: The human bladder cancer cell line T24 [American Type Culture Collection (ATCC), Manassas, VA, USA] was cultured in RMPI-1640 medium (Gibco, Invitrogen, Carlsbad, CA, USA), supplemented with 10% fetal bovine serum (FBS) (Gibco), 100 U/ml penicillin G and 100 μg/ml streptomycin.

Techniques: In Vivo, Injection, Staining

Journal: Cells

Article Title: ALG3 Promotes Peritoneal Metastasis of Ovarian Cancer through Increasing Interaction of α1,3-mannosylated uPAR and ADAM8.

doi: 10.3390/cells11193141

Figure Lengend Snippet: Figure 3. ALG3 promotes cancer stemness and proliferation in ovarian cancer cells. (A,B) Western blot analysis of ALG3 expression and Lectin blot analysis of α1,3-mannosylation in A2780 (A) and SKOV3 (B) cells after transfection with scramble siRNA, ALG3 siRNA, vector and ALG3 cDNA. (C,D) Western blot detected changes in Ki67, PCNA, cyclin E1, p21 and p27 in A2780 (C) and SKOV3 (D) cells. (E,F) EdU assay of cell proliferation ability. (G,H) Western blot analysis of Nanog and OCT4 expression. (I,J) Representative microscopy image of sphere formation assay in A2780 (I) and SKOV3 (J) cells. The bar represents 20 µm (E,F) or 10 µm (I,J).

Article Snippet: Human ovarian cancer A2780 and SKOV3 cells (ATCC, Manassas, VA, USA) and murine ovarian cancer ID8 cells (ATCC, Manassas, VA, USA) were cultured in RPMI 1640 (Gibco, Waltham, MA, USA), and human peritoneal mesothelial cells (HPMCs) were cultured in DMEM (Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA) and 100 U/mL penicillin and streptomycin.

Techniques: Western Blot, Expressing, Transfection, Plasmid Preparation, EdU Assay, Microscopy, Tube Formation Assay

Journal: Cells

Article Title: ALG3 Promotes Peritoneal Metastasis of Ovarian Cancer through Increasing Interaction of α1,3-mannosylated uPAR and ADAM8.

doi: 10.3390/cells11193141

Figure Lengend Snippet: Figure 4. ALG3 enhances the metastasis capacity of ovarian cancer cells. (A,B) Western blot anal- ysis of EMT markers (E-cadherin, N-cadherin, Snail and Vimentin) expressed in A2780 (A) and SKOV3 (B) cells after transfection with scramble siRNA, ALG3 siRNA, vector and ALG3 cDNA. (C,D) The migration and invasion potential detected by transwell migration/Matrigel invasion assay. (E,F) The migration ability detected by scratch assay. (G,H) Representative images of fluo- rescently labeled A2780 (G) and SKOV3 (H) cells adhering to HPMCs. Bars represent 5 µm (G,H), 10 µm (C,D) and 50 µm (E,F).

Article Snippet: Human ovarian cancer A2780 and SKOV3 cells (ATCC, Manassas, VA, USA) and murine ovarian cancer ID8 cells (ATCC, Manassas, VA, USA) were cultured in RPMI 1640 (Gibco, Waltham, MA, USA), and human peritoneal mesothelial cells (HPMCs) were cultured in DMEM (Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA) and 100 U/mL penicillin and streptomycin.

Techniques: Western Blot, Transfection, Plasmid Preparation, Migration, Invasion Assay, Wound Healing Assay, Labeling

Journal: Cells

Article Title: ALG3 Promotes Peritoneal Metastasis of Ovarian Cancer through Increasing Interaction of α1,3-mannosylated uPAR and ADAM8.

doi: 10.3390/cells11193141

Figure Lengend Snippet: Figure 5. uPAR α1,3-mannosylation by ALG3 increases its interaction with ADAM8. (A) Immuno- precipitated uPAR after ALG3 siRNA and ALG3 cDNA transfection in A2780 and SKOV3 cells detected by GNA and uPA. (B) Western blot analysis of uPAR expression after uPAR cDNA and uPAR MUT cDNA transfection in A2780 and SKOV3 cells. (C) Anti-uPAR Immunoprecipitations from A2780 and SKOV3 cells co-transfected with ALG3 cDNA and uPAR cDNA or uPAR MUT (mutation) cDNA and blotted with biotinylated GNA and anti-uPA. (D) Lectin ELISA detection of glycosylated uPAR was performed in A2780 and SKOV3 cells. (E) Heatmap analysis of 50 genes correlated with uPAR in GSE18520. (F) Volcano plot of differentially expressed genes between normal ovary and ovarian cancer in GSE18520. (G) Correlation analysis of uPAR and ADAM8 in GSE18520. (H) Confocal immunofluorescence shows the location of uPAR and ADAM8. The bar represents 5 µm. (I) The binding mode of the complex of ADAM8 with uPAR. Green: uPAR. Bright blue: ADAM8. (J) Immunoprecipitation shows the binding ability of uPAR and ADAM8. * p < 0.05, ** p < 0.01.

Article Snippet: Human ovarian cancer A2780 and SKOV3 cells (ATCC, Manassas, VA, USA) and murine ovarian cancer ID8 cells (ATCC, Manassas, VA, USA) were cultured in RPMI 1640 (Gibco, Waltham, MA, USA), and human peritoneal mesothelial cells (HPMCs) were cultured in DMEM (Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA) and 100 U/mL penicillin and streptomycin.

Techniques: Transfection, Western Blot, Expressing, Mutagenesis, Enzyme-linked Immunosorbent Assay, Binding Assay, Immunoprecipitation

Journal: Cells

Article Title: ALG3 Promotes Peritoneal Metastasis of Ovarian Cancer through Increasing Interaction of α1,3-mannosylated uPAR and ADAM8.

doi: 10.3390/cells11193141

Figure Lengend Snippet: Figure 6. α1,3-mannosylated uPAR binding with ADAM8 stimulates ovarian metastasis by activating the Ras/ERK signaling pathway. (A,B) Western blot analysis of ADAM8 expression after ALG3 cDNA with uPAR cDNA or uPAR MUT cDNA co-transfection and antibody blockade of ADAM8 in A2780 (A) and SKOV3 (B) cells. (C,D) Western blot analysis of Ras, p-ERK and ERK expression, as well as EMT markers (E-cadherin, N-cadherin and Vimentin) expression. (E) The migration and invasion potential detected by transwell migration/Matrigel invasion assay. (F) Representative images of fluorescently labeled A2780 and SKOV3 cells adhering to HPMCs. Bars represent 5 µm (F) and 10 µm (E).

Article Snippet: Human ovarian cancer A2780 and SKOV3 cells (ATCC, Manassas, VA, USA) and murine ovarian cancer ID8 cells (ATCC, Manassas, VA, USA) were cultured in RPMI 1640 (Gibco, Waltham, MA, USA), and human peritoneal mesothelial cells (HPMCs) were cultured in DMEM (Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA) and 100 U/mL penicillin and streptomycin.

Techniques: Binding Assay, Western Blot, Expressing, Cotransfection, Migration, Invasion Assay, Labeling

Journal: Cells

Article Title: ALG3 Promotes Peritoneal Metastasis of Ovarian Cancer through Increasing Interaction of α1,3-mannosylated uPAR and ADAM8.

doi: 10.3390/cells11193141

Figure Lengend Snippet: Figure 7. Downregulation of α1,3-mannosylation inhibits tumor peritoneal metastasis in a mouse model. (A) Ultrasound image to detect the amount of ascites (red arrow) in the peritoneal cavity of different groups (OC, si-ALG3 and GNA-). (B) Statistical analysis of the volume of ascites among the groups before mice were sacrificed. (C) Representative images of the cancer metastasis nodes (red arrows) in the peritoneum. (D) The HE-stained peritoneum was dissected longitudinally. The red arrow indicates metastatic tumor nodes. (E) Western blot analysis of EMT and stemness marker expression in peritoneal cancer tissues. (F,G) Western blots showing the expression of EMT and stemness markers in A2780 and SKOV3 cells treated with ascites from the mouse peritoneal cavity. (H) The migration and invasion potential detected by transwell migration/Matrigel invasion assay. (I) Representative images of fluorescently labeled A2780 and SKOV3 cells adhering to HPMCs. Bars represent 5 µm (I), 10 µm (H) and 100 µm (D). *** p < 0.001.

Article Snippet: Human ovarian cancer A2780 and SKOV3 cells (ATCC, Manassas, VA, USA) and murine ovarian cancer ID8 cells (ATCC, Manassas, VA, USA) were cultured in RPMI 1640 (Gibco, Waltham, MA, USA), and human peritoneal mesothelial cells (HPMCs) were cultured in DMEM (Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA) and 100 U/mL penicillin and streptomycin.

Techniques: Staining, Western Blot, Marker, Expressing, Migration, Invasion Assay, Labeling

Journal: Cancers

Article Title: PPIP5K2 Facilitates Proliferation and Metastasis of Non-Small Lung Cancer (NSCLC) through AKT Signaling Pathway

doi: 10.3390/cancers16030590

Figure Lengend Snippet: PPIP5K2 is highly expressed in NSCLC. ( A ) The distribution of PPIP5K2 gene expression in NSCLC and normal tissues in the TCGA database. ( B ) The expression distribution of PPIP5K2 gene in NSCLC tumor tissues and paired paraneoplastic tissues in the GEO database. ( C ) RT-qPCR analysis showing PPIP5K2 expression distribution cancer and paracancer tissues in 77 paired NSCLC patients. p values were determined using the Wilcox test in ( A – C ). * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Cell culture, The human NSCLC cell lines A549, NCI-H1299, NCI-H1975, and NCI-H1703 were all obtained from the American Type Culture Collection (ATCC).

Techniques: Gene Expression, Expressing, Quantitative RT-PCR

Journal: Cancers

Article Title: PPIP5K2 Facilitates Proliferation and Metastasis of Non-Small Lung Cancer (NSCLC) through AKT Signaling Pathway

doi: 10.3390/cancers16030590

Figure Lengend Snippet: PPIP5K2 promotes NSCLC cell proliferation in vitro. ( A , B ) Western blot analysis showing the knockdown and overexpression efficiency of PPIP5K2. ( C , D ) RT-qPCR analysis showing the knockdown and overexpression efficiency of PPIP5K2. ( E , F ) CCK-8 assay showing the cell proliferation when PPIP5K2 changes. ( G , H ) Cell clone formation assay showing the cell proliferation when PPIP5K2 changes. Scale bar, 100 μm. All data represent the mean ± SD of three independent experiments. p values were determined using two-way ANOVA in ( C – H ). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Cell culture, The human NSCLC cell lines A549, NCI-H1299, NCI-H1975, and NCI-H1703 were all obtained from the American Type Culture Collection (ATCC).

Techniques: In Vitro, Western Blot, Knockdown, Over Expression, Quantitative RT-PCR, CCK-8 Assay, Tube Formation Assay

Journal: Cancers

Article Title: PPIP5K2 Facilitates Proliferation and Metastasis of Non-Small Lung Cancer (NSCLC) through AKT Signaling Pathway

doi: 10.3390/cancers16030590

Figure Lengend Snippet: PPIP5K2 enhances NSCLC cells’ oncogenic capacities in vivo. ( A ) Representative images and weight of subcutaneous tumors in nude mice with injection of A549 cells. ( B ) The volume of subcutaneous tumors. ( C ) H&E staining of tumors from different groups. Scale bar, 100 μm. Data represent the mean ± SD in ( A , B ). p values were determined using Student’s t test in ( A ) and two-way ANOVA in ( B ). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Cell culture, The human NSCLC cell lines A549, NCI-H1299, NCI-H1975, and NCI-H1703 were all obtained from the American Type Culture Collection (ATCC).

Techniques: In Vivo, Injection, Staining

Journal: Cancers

Article Title: PPIP5K2 Facilitates Proliferation and Metastasis of Non-Small Lung Cancer (NSCLC) through AKT Signaling Pathway

doi: 10.3390/cancers16030590

Figure Lengend Snippet: PPIP5K2 is essential for EMT-associated cell migration in NSCLC. ( A , B ) Transwell assay showing migration of knockdown and overexpression PPIP5K2 NSCLC cells. Scale bar, 100 μm. ( C – F ) Wound healing assay showing the migration of NSCLC cells after knockdown and overexpression PPIP5K2. Scale bar, 100 μm. ( G , H ) Representative qPCR analysis showing the levels of snail1, slug, and zeb1 when knockdown PPIP5K2 in NSCLC cells. ( I , J ) Western blot analysis of E-cadherin, N-cadherin, β-catenin, Vimentin expression in PPIP5K2 knockdown (A549, H1299) and overexpression (H1975, H1703) cell lines. Data are shown as the mean ± SD of three independent experiments. p values were determined using two-way ANOVA in (A-H). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001, ns, nonsignificant.

Article Snippet: Cell culture, The human NSCLC cell lines A549, NCI-H1299, NCI-H1975, and NCI-H1703 were all obtained from the American Type Culture Collection (ATCC).

Techniques: Migration, Transwell Assay, Knockdown, Over Expression, Wound Healing Assay, Western Blot, Expressing

Journal: Cancers

Article Title: PPIP5K2 Facilitates Proliferation and Metastasis of Non-Small Lung Cancer (NSCLC) through AKT Signaling Pathway

doi: 10.3390/cancers16030590

Figure Lengend Snippet: PPIP5K2 alterations could affect the AKT/mTOR pathway. ( A , B ) Western blot analysis of a series of AKT/mTOR pathway proteins expression in A549, H1299, H1975, and H1703 cell lines.

Article Snippet: Cell culture, The human NSCLC cell lines A549, NCI-H1299, NCI-H1975, and NCI-H1703 were all obtained from the American Type Culture Collection (ATCC).

Techniques: Western Blot, Expressing

Journal: Cancers

Article Title: PPIP5K2 Facilitates Proliferation and Metastasis of Non-Small Lung Cancer (NSCLC) through AKT Signaling Pathway

doi: 10.3390/cancers16030590

Figure Lengend Snippet: PPIP5K2 promotes proliferation and metastasis of NSCLC cells through AKT/mTOR pathway. ( A , B ) Colony formation assay showing the proliferation of H1299 PPIP5K2 knockdown and H1975 PPIP5K2 OE cell lines after treatment with SC79 (10 μm), MHY1485 (10 μm), MK-2206 (1 μm), and Rapamycin (1 μm). Scale bar, 100 μm. ( C , D ) Transwell assay showing the migration of H1299 PPIP5K2 knockdown and H1975 PPIP5K2 OE cell lines after treatment with SC79 (10 μm), MHY1485 (10 μm), MK-2206 (1 μm), and Rapamycin (1 μm). Scale bar, 100 μm. All data represent the mean ± SD of three independent experiments. p values were determined using Student’s t tests in ( A – D ). ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Cell culture, The human NSCLC cell lines A549, NCI-H1299, NCI-H1975, and NCI-H1703 were all obtained from the American Type Culture Collection (ATCC).

Techniques: Colony Assay, Knockdown, Transwell Assay, Migration

Journal: Oncogene

Article Title: Aberrant NSUN2-mediated m 5 C modification of H19 lncRNA is associated with poor differentiation of hepatocellular carcinoma

doi: 10.1038/s41388-020-01475-w

Figure Lengend Snippet: a Genotyping of NSUN2-deficient HepG2 cells based on the PCR method. Because of the insertion of Neo and Puro into the double allele, two longer bands could be amplified in NSUN2-deficient cells compared with normal cells. b NSUN2 mRNA level in HepG2 and NSUN2-deficient HepG2 cells were assessed by real-time qPCR. Data are represented as the mean ± SEM ( n = 3 independent experiments). c NSUN2 protein level in HepG2 and NSUN2-deficient HepG2 cells were assessed by western blotting. Three independent experiments were performed for quantification (mean ± SEM). d Cell proliferation was determined by CCK-8 assay. Data are represented as the mean ± SEM ( n = 6 independent experiments). e Representative picture of the colony formation assay result. Three independent experiments were performed for quantification (mean ± SEM). f FACS analysis was subjected to assess the cell-cycle distribution. OD optical density; HepG2-NKD, NSUN2-deficient HepG2 cells. P values were calculated by Student’s t test. * p < 0.05, ** p < 0.01.

Article Snippet: The human hepatocellular carcinoma cell line HepG2 was purchased from ATCC (HB-8065).

Techniques: Amplification, Western Blot, CCK-8 Assay, Colony Assay

Journal: Oncogene

Article Title: Aberrant NSUN2-mediated m 5 C modification of H19 lncRNA is associated with poor differentiation of hepatocellular carcinoma

doi: 10.1038/s41388-020-01475-w

Figure Lengend Snippet: a Cell migration was determined by wound healing assay. b Trans-well invasion assay. Three independent experiments were performed for quantification (mean ± SEM). c Tubule formation assay was used to test the effect of NSUN2 deficiency on angiogenesis ability of cells in vitro. d Xenograft nude mouse model was inoculated with NSUN2-deficient HepG2 cells or the normal HepG2 cells ( n = 6/group), at day 24 after inoculation. Three independent experiments were performed for quantification (mean ± SEM). HepG2-NKD, NSUN2-deficient HepG2 cells. The data are represented as mean ± SEM, obtained in three independent experiments. P values were calculated by Student’s t test. * p < 0.05, ** p < 0.01.

Article Snippet: The human hepatocellular carcinoma cell line HepG2 was purchased from ATCC (HB-8065).

Techniques: Migration, Wound Healing Assay, Invasion Assay, Tube Formation Assay, In Vitro

Journal: Oncogene

Article Title: Aberrant NSUN2-mediated m 5 C modification of H19 lncRNA is associated with poor differentiation of hepatocellular carcinoma

doi: 10.1038/s41388-020-01475-w

Figure Lengend Snippet: a Volcano plot of differentially expressed genes (DEGs) after NSUN2 depletion. b Heat-map expression profile of 2681 significantly dysregulated RNAs between HepG2 and NSUN2-deficient HepG2 cells. c Statistical analysis of KEGG pathway enrichment for the DEGs. d Histogram showed the number of total m 5 C sites and annotated RNAs in HepG2 cells and NSUN2-deficient HepG2 cells. e Statistical analysis of KEGG pathway enrichment for the differentially methylated genes (DMGs).

Article Snippet: The human hepatocellular carcinoma cell line HepG2 was purchased from ATCC (HB-8065).

Techniques: Expressing, Methylation

Journal: Oncogene

Article Title: Aberrant NSUN2-mediated m 5 C modification of H19 lncRNA is associated with poor differentiation of hepatocellular carcinoma

doi: 10.1038/s41388-020-01475-w

Figure Lengend Snippet: a Deficiency of NSUN2 decreased the level of H19 RNA in HepG2 cells, while restore NSUN2 could recover the level of H19 RNA in NSUN2-deficient HepG2 cells. b Sanger-based validation of m 5 C sites within H19 RNA. cDNA was amplified by PCR using specific primers for bisulfite-treated RNAs. c Quantitative determination of methylation level of H19 RNA C986 site in normal and NSUN2-deficient HepG2 cells through pyrosequencing. d Pyrosequencing result demonstrated restore of NSUN2 could recover the methylation level of H19 C986 site in NSUN2-deficient HepG2 cells. Mock, cells transfected with empty pcDNA3.1 vector; pNSUN2, cells transfected with pcDNA3.1-NSUN2 vector; HepG2-NKD, NSUN2-deficient HepG2 cells. The results are expressed as the mean ± SEM from three independent experiments, ** p < 0.01, *** p < 0.001.

Article Snippet: The human hepatocellular carcinoma cell line HepG2 was purchased from ATCC (HB-8065).

Techniques: Biomarker Discovery, Amplification, Methylation, Transfection, Plasmid Preparation

Journal: Oncogene

Article Title: Aberrant NSUN2-mediated m 5 C modification of H19 lncRNA is associated with poor differentiation of hepatocellular carcinoma

doi: 10.1038/s41388-020-01475-w

Figure Lengend Snippet: a NSUN2 deficiency decreased the half-life of H19 RNA in HepG2 cells. Cells were exposed to actinomycin D (5 μg/ml), whereupon the cellular RNA was isolated at times indicated. Real-time qPCR against GAPDH was performed to assess the half-lives of H19 RNA and β-actin mRNA b Restore of NSUN2 could recover the half-life of H19 RNA at a certain level in NSUN2-deficient HepG2 cells. Three independent experiments were performed for quantification (mean ± SEM). c HepG2 cells or NSUN2-deficient HepG2 cells were transfected with plasmids pimrGLO-H19-wt or pimrGLO-H19-mut. After 48 h, Firefly luciferase activity was determined and normalized against Renilla luciferase activity. Data are represented as the mean ± SEM from three independent experiments. d H19 expression in 55 pairs of liver cancer tissues and matched no-cancerous liver tissues. The relative expression levels were normalized to the expression amount of the GAPDH gene. e H19 RNA methylation status in 55 pairs of liver cancer tissues and matched no-cancerous liver tissues. f Pearson’s correlation analysis demonstrating that H19 expression is positively correlated with the m 5 C level in 110 samples (55 liver cancer tissues and 55 matched no-cancerous liver tissues). g NSUN2 mRNA level in 55 pairs of liver cancer tissues and matched no-cancerous liver tissues. h NSUN2 mRNA level in 16 pairs of liver cancer tissues and matched no-cancerous liver tissues with H19 C986 site methylation level difference over 10%. The relative expression levels were normalized to the expression amount of the GAPDH gene. mock, cells transfected with empty pcDNA3.1 vector; pNSUN2, cells transfected with pcDNA3.1-NSUN2 vector; HepG2-NKD, NSUN2-deficient HepG2 cells. * p < 0.05, ** p < 0.01.

Article Snippet: The human hepatocellular carcinoma cell line HepG2 was purchased from ATCC (HB-8065).

Techniques: Isolation, Transfection, Luciferase, Activity Assay, Expressing, Methylation, Plasmid Preparation

Journal: Oncogene

Article Title: Aberrant NSUN2-mediated m 5 C modification of H19 lncRNA is associated with poor differentiation of hepatocellular carcinoma

doi: 10.1038/s41388-020-01475-w

Figure Lengend Snippet: a Proteins retrieved by H19 probes visualized by silver staining. b G3BP1 protein level in HepG2 and NSUN2-deficient HepG2 cells were assessed by western blotting. c RIP was performed using HepG2 or NSUN2-deficient HepG2 cell lysate and either anti-G3BP1 or Normal Rabbit IgG as the immunoprecipitating antibody. Purified RNA was analyzed by real-time qPCR or RT-PCR using primers specific for H19 or GAPDH . Data are represented as means ± SEM from three independent experiments. d In vitro protein-RNA-binding assays (rEMSA) were performed using H19 oligonucleotides with or without m 5 C as well as the indicated amounts of purified G3BP1 protein (200–400 ng/reaction). HepG2-NKD, NSUN2-deficient HepG2 cells.

Article Snippet: The human hepatocellular carcinoma cell line HepG2 was purchased from ATCC (HB-8065).

Techniques: Silver Staining, Western Blot, Purification, Reverse Transcription Polymerase Chain Reaction, In Vitro, RNA Binding Assay

Journal: ACS Omega

Article Title: MiR-199 Reverses the Resistance to Gemcitabine in Pancreatic Cancer by Suppressing Stemness through Regulating the Epithelial–Mesenchymal Transition

doi: 10.1021/acsomega.1c02945

Figure Lengend Snippet: GEM-resistant SW1990 cells (SW1990/GZ cells) were successfully established. (A) Cell viability of SW1990 and SW1990/GZ cells was detected by the CCK-8 assay, and the IC50 values were calculated. (B) Cell sphere formation rate was determined using the sphere formation assay. (C) Number of spheres was counted in the colony formation assay. (D) Flow cytometry was used to measure the apoptosis. (E) Transwell assay was used to determine the migration ability. (F) Expression level of E-cadherin, Vimentin, and β-catenin was determined by the Western blotting assay (* p < 0.05 vs SW1990, ** p < 0.01 vs SW1990).

Article Snippet: The human pancreatic cancer cell line SW1990 cells were purchased from ATCC (Maryland, USA) and cultured in the Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) at 37 °C and 5% CO 2 .

Techniques: CCK-8 Assay, Tube Formation Assay, Colony Assay, Flow Cytometry, Transwell Assay, Migration, Expressing, Western Blot

Journal: ACS Omega

Article Title: MiR-199 Reverses the Resistance to Gemcitabine in Pancreatic Cancer by Suppressing Stemness through Regulating the Epithelial–Mesenchymal Transition

doi: 10.1021/acsomega.1c02945

Figure Lengend Snippet: Self-renewal of SW1990/GZ cells was suppressed by miR-199. (A) Morphology of cells was observed under an inverted microscope. (B) Cell sphere formation rate was determined using the sphere formation assay. (C) Number of spheres was counted in the colony formation assay. (D) Expression of CD44, ALDH1, and Nanog was determined by the Western blotting assay (** p < 0.01 vs miR-NC + SW1990/GZ).

Article Snippet: The human pancreatic cancer cell line SW1990 cells were purchased from ATCC (Maryland, USA) and cultured in the Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) at 37 °C and 5% CO 2 .

Techniques: Inverted Microscopy, Tube Formation Assay, Colony Assay, Expressing, Western Blot

Journal: ACS Omega

Article Title: MiR-199 Reverses the Resistance to Gemcitabine in Pancreatic Cancer by Suppressing Stemness through Regulating the Epithelial–Mesenchymal Transition

doi: 10.1021/acsomega.1c02945

Figure Lengend Snippet: Inhibitory effect of GEM on the proliferation and migration of SW1990/GZ cells was enhanced by miR-199. (A) Cell viability was detected by the CCK-8 assay; (B) tFlow cytometry was used to measure the apoptosis. (C) Transwell assay was used to determine the migration ability (** p < 0.01 vs miR-NC + SW1990/GZ).

Article Snippet: The human pancreatic cancer cell line SW1990 cells were purchased from ATCC (Maryland, USA) and cultured in the Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) at 37 °C and 5% CO 2 .

Techniques: Migration, CCK-8 Assay, Cytometry, Transwell Assay

Journal: ACS Omega

Article Title: MiR-199 Reverses the Resistance to Gemcitabine in Pancreatic Cancer by Suppressing Stemness through Regulating the Epithelial–Mesenchymal Transition

doi: 10.1021/acsomega.1c02945

Figure Lengend Snippet: Inhibitory effect of GEM on EMT progression of SW1990/GZ cells was enhanced by miR-199. The expression level of E-cadherin, Vimentin, and β-catenin was determined by the Western blotting assay (** p < 0.01 vs miR-NC + SW1990/GZ).

Article Snippet: The human pancreatic cancer cell line SW1990 cells were purchased from ATCC (Maryland, USA) and cultured in the Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) at 37 °C and 5% CO 2 .

Techniques: Expressing, Western Blot

Journal: ACS Omega

Article Title: MiR-199 Reverses the Resistance to Gemcitabine in Pancreatic Cancer by Suppressing Stemness through Regulating the Epithelial–Mesenchymal Transition

doi: 10.1021/acsomega.1c02945

Figure Lengend Snippet: miR-199 targeted Snail to regulate its expression in SW1990/GZ cells. (A) Binding site between miR-199 and Snail was confirmed by the dual-luciferase gene reporter assay (** p < 0.01 vs miR-NC). (B) Expression level of Snail was determined by the Western blotting assay (** p < 0.01 vs miR-NC + SW1990/GZ).

Article Snippet: The human pancreatic cancer cell line SW1990 cells were purchased from ATCC (Maryland, USA) and cultured in the Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) at 37 °C and 5% CO 2 .

Techniques: Expressing, Binding Assay, Luciferase, Reporter Assay, Western Blot

Journal: ACS Omega

Article Title: MiR-199 Reverses the Resistance to Gemcitabine in Pancreatic Cancer by Suppressing Stemness through Regulating the Epithelial–Mesenchymal Transition

doi: 10.1021/acsomega.1c02945

Figure Lengend Snippet: Inhibitory effect of miR-199 on self-renewal of SW1990/GZ cells was abolished by overexpressing Snail. (A) Transfection efficacy was verified by the Western blotting assay. (B) Cell sphere formation rate was determined using the sphere formation assay. (C) Number of spheres was counted in the colony formation assay. (D) Expression of CD44, ALDH1, and Nanog was determined by the Western blotting assay (* p < 0.05 vs SW1990/GZ, ## p < 0.01 vs GEM + SW1990/GZ, and && p < 0.01 vs miR-199 + GEM + SW1990/GZ).

Article Snippet: The human pancreatic cancer cell line SW1990 cells were purchased from ATCC (Maryland, USA) and cultured in the Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) at 37 °C and 5% CO 2 .

Techniques: Transfection, Western Blot, Tube Formation Assay, Colony Assay, Expressing

Journal: ACS Omega

Article Title: MiR-199 Reverses the Resistance to Gemcitabine in Pancreatic Cancer by Suppressing Stemness through Regulating the Epithelial–Mesenchymal Transition

doi: 10.1021/acsomega.1c02945

Figure Lengend Snippet: Enhancement of miR-199 on the inhibitory effect of GEM against the proliferation and migration ability of SW1990/GZ cells was abolished by overexpressing Snail. (A) Flow cytometry was used to measure the apoptosis. (C) Transwell assay was used to determine the migration ability (* p < 0.05 vs SW1990/GZ, ## p < 0.01 vs GEM + SW1990/GZ, & p < 0.05 vs miR-199 + GEM + SW1990/GZ, and && p < 0.01 vs miR-199 + GEM + SW1990/GZ).

Article Snippet: The human pancreatic cancer cell line SW1990 cells were purchased from ATCC (Maryland, USA) and cultured in the Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) at 37 °C and 5% CO 2 .

Techniques: Migration, Flow Cytometry, Transwell Assay

Journal: ACS Omega

Article Title: MiR-199 Reverses the Resistance to Gemcitabine in Pancreatic Cancer by Suppressing Stemness through Regulating the Epithelial–Mesenchymal Transition

doi: 10.1021/acsomega.1c02945

Figure Lengend Snippet: Enhancement of miR-199 on the inhibitory effect of GEM against the EMT progressing of SW1990/GZ cells was abolished by overexpressing Snail. The expression level of E-cadherin, Vimentin, and β-catenin was determined by the Western blotting assay (* p < 0.05 vs SW1990/GZ, ## p < 0.01 vs GEM + SW1990/GZ, & p < 0.05 vs miR-199 + GEM + SW1990/GZ, and && p < 0.01 vs miR-199 + GEM + SW1990/GZ).

Article Snippet: The human pancreatic cancer cell line SW1990 cells were purchased from ATCC (Maryland, USA) and cultured in the Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) at 37 °C and 5% CO 2 .

Techniques: Expressing, Western Blot

Journal: ACS Omega

Article Title: MiR-199 Reverses the Resistance to Gemcitabine in Pancreatic Cancer by Suppressing Stemness through Regulating the Epithelial–Mesenchymal Transition

doi: 10.1021/acsomega.1c02945

Figure Lengend Snippet: Strengthening of miR-199 on the inhibitory effect of GEM against in vivo tumor growth of SW1990/GZ cells was abolished by overexpressing Snail. (A) Tumor volume was recorded during the treatments on days 1, 4, 7, 10, 14, and 16. (B) Tumor weights were recorded at the end of the animal experiment. (C) Picture of tumors was taken at the end of the animal experiment (** p < 0.01 vs SW1990/GZ, ## p < 0.01 vs GEM + SW1990/GZ, and && p < 0.01 vs miR-199 + GEM + SW1990/GZ).

Article Snippet: The human pancreatic cancer cell line SW1990 cells were purchased from ATCC (Maryland, USA) and cultured in the Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) at 37 °C and 5% CO 2 .

Techniques: In Vivo