Journal: Journal of Cancer

Article Title: Preliminary study on SPECT/CT imaging of pancreatic cancer xenografts by targeting integrin α5 in pancreatic stellate cells

doi: 10.7150/jca.51190

Figure Lengend Snippet: Western blotting assay (A) and cell immunofluorescence staining (B) of ITGA5 expression in PSCs without or with SW1990-CM treatment. * P < 0.05 , n = 3.

Article Snippet: The human pancreatic cancer SW1990 cells and PSCs were obtained from Tongpai (Shanghai) Biotechnology Co., LTD. Anti-ITGA5 antibody and FITC-labeled secondary antibody were purchased from Beijing Bioss Biotechnology Co., LTD. DMEM and fetal bovine serum (FBS) were purchased from Gibco Life Technology Co., LTD.

Techniques: Western Blot, Immunofluorescence, Staining, Expressing

Journal: Journal of Cancer

Article Title: Preliminary study on SPECT/CT imaging of pancreatic cancer xenografts by targeting integrin α5 in pancreatic stellate cells

doi: 10.7150/jca.51190

Figure Lengend Snippet: Representative 125 I-AV3 SPECT/CT imaging at 4 h (A), corresponding SPECT/CT imaging with a blocking dose of AV3 (B) and ITGA5 IHC staining (C) of pancreatic cancer xenografts in nude mice. The xenografts tumors in nude mice were established using SW1990 cells injection alone in the left lower limb (indicated by asterisk) or SW1990 + PSCs co-injection in the right lower limb (indicated by arrow). Pancreatic cancer-bearing nude mice were injected about 1 mCi 125 I-AV3 from tail vein before the imaging study. Repeated experiments showed similar results.

Article Snippet: The human pancreatic cancer SW1990 cells and PSCs were obtained from Tongpai (Shanghai) Biotechnology Co., LTD. Anti-ITGA5 antibody and FITC-labeled secondary antibody were purchased from Beijing Bioss Biotechnology Co., LTD. DMEM and fetal bovine serum (FBS) were purchased from Gibco Life Technology Co., LTD.

Techniques: Single Photon Emission Computed Tomography, Imaging, Blocking Assay, Immunohistochemistry, Injection

Journal: American Journal of Translational Research

Article Title: Ganoderma lucidum fruiting body extracts inhibit colorectal cancer by inducing apoptosis, autophagy, and G0/G1 phase cell cycle arrest in vitro and in vivo

doi:

Figure Lengend Snippet: GLE and cisplatin inhibited the proliferation of four tumor cell lines in a dose- and time- dependent manner. Four tumor cells (A549, SW1990, SKOV3 and HCT116 cells) were treated with GLE and cisplatin for 12 h, 24 h and 48 h, and their viabilities were determined by CCK8 assay. GLE reduced the cell viability of A549 cells (A), SW1990 cells (B), SKOV3 cells (C) and HCT116 cells (D). Cisplatin reduced the cell viability of four tumor cell lines (E).

Article Snippet: A549, SW1990, SKOV3 and HCT116 were purchased from KeyGEN biotechnology company and cultured in DMEM, DMEM, DMEM and 1640 medium, respectively, containing 10% fetal bovine serum, penicillin (50 U/mL) and streptomycin (50 U/mL) in a humidified incubator with 5% CO 2 at 37°C.

Techniques: CCK-8 Assay

Journal: Frontiers in Immunology

Article Title: Design and selection of optimal ErbB-targeting bispecific antibodies in pancreatic cancer

doi: 10.3389/fimmu.2023.1168444

Figure Lengend Snippet: Pre-clinical evaluation of the three selected lead BiXAb™ 3Patri-2Trastu-Fc, 3Patri-1Matu-Fc, and 3Patri-1Cetu-Fc. (A) Tumor growth (left) and survival (middle) of BiXAb™-treated mice xenografted with Sw1990 and PDX P2846 PDAC cells. The percentage of tumor growth inhibition (TGI) at treatment end, and the 50% survival benefit (days) are indicated in the right panel. (B) Immunohistochemistry analysis of BiXAb™ penetration in Sw1990 cell xenografts. BiXAb™ were detected using peroxidase-conjugated anti-human Fc. IRR BiXAb™, BiXAb™ targeting HLA-DR and CD5 (negative control). NaCl was used as negative control. (C) Tumor penetration of 3Patri-1Cetu-Fc evaluated in whole tumor slices at lower magnification. The external (Ext) and internal (Int) parts of the tumor slice are indicated.

Article Snippet: Sw1990 cells (3x10 6 cells per mouse) or the PDX P2846 at passage 8 (P8) since surgery (125 mm 3 tumor piece per mouse) were injected subcutaneously in the right flank, or transplanted intrascapularly in the brown fat, respectively, of 6 week/old female Swiss nude (Crl : NU(Ico)-Foxn1nu) mice from Charles River (Wilmington, MA).

Techniques: Inhibition, Immunohistochemistry, Negative Control

Journal: Frontiers in Immunology

Article Title: Design and selection of optimal ErbB-targeting bispecific antibodies in pancreatic cancer

doi: 10.3389/fimmu.2023.1168444

Figure Lengend Snippet: NK cell engagement and receptor expression. (A) Immunophenotyping by flow cytometry of NK cells in Sw1990 cell xenografts resected from BiXAb™-treated mice. Dissociated cells were labeled with human CD49-, NKp46-, IFNγ- and CD107-specific antibodies. (B) Immunofluorescence analysis of EGFR expression by immunofluorescence in 3Patri-1Cetu-Fc-, IRR BiXAb™- and NaCl-treated Sw1990 cell xenografts. (C) Western blot analysis of ErbB receptor expression in Sw1990 cell xenografts from mice treated with NaCl, 3Patri-2Trastu-Fc, 3Patri-1Matu-Fc, or 3Patri-1Cetu-Fc. Tubulin was used as loading control. (D) Western blots were quantified using the LI-COR Odyssey imaging system. ErbB expression level was normalized to tubulin expression level. Each experiment was performed using three Sw1990 cell xenografted mice. IRR BiXAb™, negative control. Data are the mean ± SEM. P-values: p <0.05 (*), p <0.01 (**), p <0.001 (***), ns (not significant).

Article Snippet: Sw1990 cells (3x10 6 cells per mouse) or the PDX P2846 at passage 8 (P8) since surgery (125 mm 3 tumor piece per mouse) were injected subcutaneously in the right flank, or transplanted intrascapularly in the brown fat, respectively, of 6 week/old female Swiss nude (Crl : NU(Ico)-Foxn1nu) mice from Charles River (Wilmington, MA).

Techniques: Expressing, Flow Cytometry, Labeling, Immunofluorescence, Western Blot, Control, Imaging, Negative Control

Journal: Frontiers in Immunology

Article Title: Design and selection of optimal ErbB-targeting bispecific antibodies in pancreatic cancer

doi: 10.3389/fimmu.2023.1168444

Figure Lengend Snippet: In vivo effects of the best-in-class BiXAb™ 3Patri-1Cetu-Fc versus 2MAbs and 1MAbs. Tumor growth (left panel) and survival (middle panel) of mice xenografted with Sw1990 and PDX P2846 PDAC cells treated with the indicated antibodies (n=10 mice/group). The percentage of tumor growth inhibition (TGI), and the 50% survival benefit (days) are indicated in the right panel. The BiXAb™ 3Patri-1Cetu-Fc was used at Fab equimolarity (8.5 mg/kg) or Fc equimolarity (17 mg/kg).

Article Snippet: Sw1990 cells (3x10 6 cells per mouse) or the PDX P2846 at passage 8 (P8) since surgery (125 mm 3 tumor piece per mouse) were injected subcutaneously in the right flank, or transplanted intrascapularly in the brown fat, respectively, of 6 week/old female Swiss nude (Crl : NU(Ico)-Foxn1nu) mice from Charles River (Wilmington, MA).

Techniques: In Vivo, Inhibition

Journal: Frontiers in Immunology

Article Title: Design and selection of optimal ErbB-targeting bispecific antibodies in pancreatic cancer

doi: 10.3389/fimmu.2023.1168444

Figure Lengend Snippet: Characterization of 3Patri-1Cetu-Fc tumor penetration, and its effects on tumor angiogenesis and proliferation in mice xenografted with Sw1990 cells. (A) Immunohistochemistry analysis of the penetration of the BiXAb™ 3Patri-1Cetu-Fc, the 2MAbs 3Patri+1Cetu, and 3Patri and 1Cetu in tumor xenograft sections (x20 magnification; left panels). Antibodies were detected with a peroxidase-conjugated anti-human Fc. Human Fc-positive cells (right panel) were quantified in the whole tumor section using the QuPath software. Immunohistochemistry analysis (x20 magnification; left panel) of CD31 angiogenesis (B) and Ki67 proliferation (C) in Sw1990 cell xenografts. CD31+ microvessel density was estimated as the mean value in ten histological fields using the Image J software. The percentage of Ki-67+ cells in tumors was quantified in the whole tumor section using the Qupath software. IRR BiXAb™, negative control BiXAb™. Data are the mean ± SEM (n=5 mice/group). P-values: p <0.05 (*), p <0.01 (**), p <0.001 (***), ns (not significant). Immunohistochemistry analysis of P2846 xenografts is in Supplementary Figure 6 .

Article Snippet: Sw1990 cells (3x10 6 cells per mouse) or the PDX P2846 at passage 8 (P8) since surgery (125 mm 3 tumor piece per mouse) were injected subcutaneously in the right flank, or transplanted intrascapularly in the brown fat, respectively, of 6 week/old female Swiss nude (Crl : NU(Ico)-Foxn1nu) mice from Charles River (Wilmington, MA).

Techniques: Immunohistochemistry, Software, Negative Control

Journal: Frontiers in Pharmacology

Article Title: Phytopharmacology and Clinical Updates of Berberis Species Against Diabetes and Other Metabolic Diseases

doi: 10.3389/fphar.2020.00041

Figure Lengend Snippet: In vitro activity of extracts and/or isolated compounds from Berberis species against diabetes and metabolic diseases.

Article Snippet: Berberine (BBR) , CEM, HCT-116, HepG2.2.15, SW1990, HT1080 and 293T cell lines , ↑gene expression of the insulin receptor , ( ) .

Techniques: In Vitro, Activity Assay, Isolation, Binding Assay, Activation Assay, Cell Culture, Expressing, Small Interfering RNA, Blocking Assay, Gene Expression, Inhibition, Translocation Assay, Phospho-proteomics, Modification

Journal: Cell Death & Disease

Article Title: MiR-30a regulates cancer cell response to chemotherapy through SNAI1/IRS1/AKT pathway

doi: 10.1038/s41419-019-1326-6

Figure Lengend Snippet: a , b Drug-resistance cell line was established as described in the Materials and methods. Gemcitabine sensitivity of both SW1990-R cells and its parental cells ( a ) were tested by MTS assay and the IC50 values were calculated ( b ). * P < 0.05 and ** P < 0.01, compared with control conditions. c Sequence length distribution in SW1990-R and SW1990 cells. d Summary of sequencing reads mapped to various types of sRNA in SW1990-R and SW1990 cells. e Cluster analysis of changed miRNAs in SW1990-R cells compared with SW1990 cells. A line indicates a microRNA gene, and a column indicates a sample. Two samples were contained in each group. Both upregulated (red) and downregulated miRNAs (blue) were identified. f Expression s of ten identified miRNAs were detected by qRT-PCR to validate microarray data. * P < 0.05, compared with SW1990. Points, mean values for three independent experiments; error bars, ±SEM

Article Snippet: Capan-2 and MIA PaCa-2 were purchased from Obio Technology (Shanghai, China) Corp., Ltd. SW1990, PANC-1 and MIA PaCa-2 were cultured in DMEM, Capan-2 was maintained in McCoy’s 5A and BxPC-3 was cultured in RMPI 1640, all supplemented with 10% fetal bovine serum, 100 U/ml of penicillin and 100 μg/ml of streptomycin.

Techniques: MTS Assay, Control, Sequencing, Expressing, Quantitative RT-PCR, Microarray

Journal: Cell Death & Disease

Article Title: MiR-30a regulates cancer cell response to chemotherapy through SNAI1/IRS1/AKT pathway

doi: 10.1038/s41419-019-1326-6

Figure Lengend Snippet: a – d GO analysis of dysregulated miRNA-mRNAs between SW1990-R and SW1990 cells. The significantly enriched biological process ( a ), cellular component ( b ), and molecular function ( c ) were showed by Directed Acyclic Graph (DAG). GO terms in box are enriched ones while those in circle are non-enriched parents. Color depth represents the enrichment degree. The number of dysregulated genes in each significantly enriched GO term was displayed as histogram ( d ). e Analysis of top 20 overrepresented KEGG pathways between SW1990-R and SW1990 cell line. The size of each circle stands for the number of dysregulated genes enriched in corresponding pathway. The rich factor was calculated using the number of enriched genes divided by the number of all background genes in corresponding pathway. Q -value was calculated using the Benjamini–Hochberg correction. When the number of genes is greater, the rich factor is greater and the Q -value is closer to zero, then the enrichment is more significant

Article Snippet: Capan-2 and MIA PaCa-2 were purchased from Obio Technology (Shanghai, China) Corp., Ltd. SW1990, PANC-1 and MIA PaCa-2 were cultured in DMEM, Capan-2 was maintained in McCoy’s 5A and BxPC-3 was cultured in RMPI 1640, all supplemented with 10% fetal bovine serum, 100 U/ml of penicillin and 100 μg/ml of streptomycin.

Techniques:

Journal: Cell Death & Disease

Article Title: MiR-30a regulates cancer cell response to chemotherapy through SNAI1/IRS1/AKT pathway

doi: 10.1038/s41419-019-1326-6

Figure Lengend Snippet: a , b SW1990 cells were transfected with miR-30a precursor (miR-30a) or scramble control lentivirus (miR-Con). MiR-30a level was confirmed by qRT-PCR ( a ). Gemcitabine sensitivity was determined by MTS assays ( b ). * P < 0.05 and ** P < 0.01, compared with miR-Con. c , d SW1990 cells were transfected with miR-30a inhibitor or scramble control (miR-Con). MiR-30a expression was detected by qRT-PCR ( c ). Gemcitabine sensitivity was determined by MTS assays ( d ). * P < 0.05 and ** P < 0.01, compared with miR-Con. e SW1990 cells were transfected with indicated constructs and then treated with 2 μM gemcitabine for 72 h. The colony forming assay was performed. Representative micrographs (left) and quantification (right) of crystal violet-stained cell colonies were displayed. f SW1990 cells were transfected with indicated constructs. Cells were then treated and detected as in e . g , h SW1990 cells were transfected with miR-30a or miR-Con. Cell growth were determined by MTS assays ( g ). Representative micrographs (left) of colony formation, as well as quantification (right) of crystal violet-stained cell colonies were displayed ( h ). n = 3 wells per group. * P < 0.05 and ** P < 0.01, compared with miR-Con. i , j SW1990 cells were transfected with miR-30a inhibitor or miR-Con. Cell growth and colony formation assay were determined as in g and h , respectively. n = 3 wells per group. * P < 0.05 and ** P < 0.01, compared with miR-Con. k , l SW1990-R cells were transfected with miR-30a precursor (miR-30a) or scramble control lentivirus (miR-Con). MiR-30a level was confirmed by qRT-PCR ( k ). Gemcitabine sensitivity was determined by MTS assays ( l ). * P < 0.05 and ** P < 0.01, compared with miR-Con. m SW1990-R cells after transfection with indicated constructs were treated with 6uM gemcitabine for 72 h. The colony forming assay was performed as in e . Points, mean values for three independent experiments; error bars, ±SEM

Article Snippet: Capan-2 and MIA PaCa-2 were purchased from Obio Technology (Shanghai, China) Corp., Ltd. SW1990, PANC-1 and MIA PaCa-2 were cultured in DMEM, Capan-2 was maintained in McCoy’s 5A and BxPC-3 was cultured in RMPI 1640, all supplemented with 10% fetal bovine serum, 100 U/ml of penicillin and 100 μg/ml of streptomycin.

Techniques: Transfection, Control, Quantitative RT-PCR, Expressing, Construct, Staining, Colony Assay

Journal: Cell Death & Disease

Article Title: MiR-30a regulates cancer cell response to chemotherapy through SNAI1/IRS1/AKT pathway

doi: 10.1038/s41419-019-1326-6

Figure Lengend Snippet: a The effect of miR-30a modulation (miR-con, miR-30a and miR-30a inhibitor) on IRS1 and SNAI1 protein expression by western blot analysis in SW1990 cells. b MiR-30a targets SNAI1 directly. Representative seed sequences for miR-30a on the SNAI1 3′UTR was shown (left). 293T cells were transiently transfected with pmirGlo plasmids encoding wild-type or mutated 3′UTR sequences of SNAI1, and oligos. Luciferase activities were then measured 24 h after transfection (right). Firefly luciferase was normalized to Renilla luciferase. The data are represented as mean ± SEM ( n = 3). c – e Knockdown of SNAI1 by transfection of two different SNAI1 siRNAs (si-SNAI1–1, si -SNAI1–2) in SW1990 cells. Cell growth ( c ) and gemcitabine sensitivity ( d ) were tested by MTS assay. Downstream proteins of SNAI1 were examined by western blot ( e ). * P < 0.05 and ** P < 0.01, compared with si-NC. f – h Indicated constructs (miR-Con, miR-30a, and/or SNAI1) were transfected in SW1990 cells. Cell growth ( f ) and gemcitabine sensitivity ( g ) were tested by MTS assay. Downstream proteins of SNAI1 were examined by western blot ( h ). * P < 0.05 and ** P < 0.01, compared with miR-Con. # P < 0.05 and ## P < 0.01, compared with miR-30a. Points, mean values for three independent experiments; error bars, ±SEM

Article Snippet: Capan-2 and MIA PaCa-2 were purchased from Obio Technology (Shanghai, China) Corp., Ltd. SW1990, PANC-1 and MIA PaCa-2 were cultured in DMEM, Capan-2 was maintained in McCoy’s 5A and BxPC-3 was cultured in RMPI 1640, all supplemented with 10% fetal bovine serum, 100 U/ml of penicillin and 100 μg/ml of streptomycin.

Techniques: Expressing, Western Blot, Transfection, Luciferase, Knockdown, MTS Assay, Construct

Journal: Cell Death & Disease

Article Title: MiR-30a regulates cancer cell response to chemotherapy through SNAI1/IRS1/AKT pathway

doi: 10.1038/s41419-019-1326-6

Figure Lengend Snippet: a , b SW1990 cells were transfected with indicated constructs. FOXO3a expression in SW1990 cells with or without miR-30a overexpression was detected by qRT-PCR ( a ). * P < 0.05, compared with control conditions. Protein expression of phospho-FOXO3a and total FOXO3a level were detected by western blot ( b ). c MiR-30a expression in SW1990-R cells and its parental cells was detected by qRT-PCR. * P < 0.05, compared with SW1990. d Gemcitabine-induced changes in protein expression between SW1990 cells and its parental cells were examined by western blot. e Computational prediction of FOXO3a binding site (S1) on 5’ miR-30a regulatory region. f Representative anti-FoxO3a ChIP-PCR showing FoxO3a binding to the miR-30a promoter. g Knockdown of FOXO3a with two different FOXO3a siRNAs (si-FOXO3a-1, si-FOXO3a-2) in SW1990 cells. MiR-30a level was determined by qRT-PCR subsequently. * P < 0.05, compared with control conditions. h , i SW1990 cells were treated with or without 10% FBS after overnight serum starvation. The P-AKT, P-ERK, P-FOXO3a, and FOXO3a in cell lysates were tested by western blot ( h ); mRNA level of FOXO3a and miR-30a were detected by qRT-PCR ( i ). * P < 0.05, compared with FBS group. j Transient knockdown of IRS1 with siRNA in SW1990 cells. MiR-30a and FOXO3a level were detected by qRT-PCR subsequently. * P < 0.05, compared with control conditions. Points, mean values for three independent experiments; error bars, ±SEM

Article Snippet: Capan-2 and MIA PaCa-2 were purchased from Obio Technology (Shanghai, China) Corp., Ltd. SW1990, PANC-1 and MIA PaCa-2 were cultured in DMEM, Capan-2 was maintained in McCoy’s 5A and BxPC-3 was cultured in RMPI 1640, all supplemented with 10% fetal bovine serum, 100 U/ml of penicillin and 100 μg/ml of streptomycin.

Techniques: Transfection, Construct, Expressing, Over Expression, Quantitative RT-PCR, Control, Western Blot, Binding Assay, Knockdown