Journal: Cancer Management and Research

Article Title: LncRNA SLC16A1-AS1 Suppresses Cell Proliferation in Cervical Squamous Cell Carcinoma (CSCC) Through the miR-194/SOCS2 Axis

doi: 10.2147/CMAR.S276629

Figure Lengend Snippet: Downregulation of SLC16A1-AS1 predicted poor survival of CSCC patients. ( A ) The expression of SLC16A1-AS1 in CSCC and paired non-tumor tissues from CSCC patients (n = 60) were determined by RT-qPCR. PCRs were performed in triplicate and mean values were presented and compared. *** p < 0.05. ( B ) The 60 patients were divided into high and low SLC16A1-AS1 level groups (n = 30, with median expression level of SLC16A1-AS1 in CSCC tissues as cutoff value) and survival curves were plotted.

Article Snippet: Human CSCC cell line SiHa (ATCC, USA) was used.

Techniques: Expressing, Quantitative RT-PCR

Journal: Cancer Management and Research

Article Title: LncRNA SLC16A1-AS1 Suppresses Cell Proliferation in Cervical Squamous Cell Carcinoma (CSCC) Through the miR-194/SOCS2 Axis

doi: 10.2147/CMAR.S276629

Figure Lengend Snippet: SLC16A1-AS1 interacted with miR-194. ( A ) Schematic model of interaction between SLC16A1-AS1 and miR-194. ( B ) Luciferase reporter assay for luciferase activity of SLC16A1-AS1-reporter with mutated miR-194 binding sites in SiHa cells co-transfected with miR-194 mimics. n = 3. ** p < 0.01.

Article Snippet: Human CSCC cell line SiHa (ATCC, USA) was used.

Techniques: Luciferase, Reporter Assay, Activity Assay, Binding Assay, Transfection

Journal: Cancer Management and Research

Article Title: LncRNA SLC16A1-AS1 Suppresses Cell Proliferation in Cervical Squamous Cell Carcinoma (CSCC) Through the miR-194/SOCS2 Axis

doi: 10.2147/CMAR.S276629

Figure Lengend Snippet: Overexpression of SLC16A1-AS1 and miR-194 did not affect the expression of each other. ( A and B ) The effects of overexpression of SLC16A1-AS1 and miR-194 mimic transfection on SLC16A1-AS1 and miR-194 expression in siHa cells. n = 3. ( C and D ) The effects of SLC16A1-AS1 overexpression and miR-194 mimic transfection on miR-194 and SLC16A1-AS1 expression in SiHa cells. n = 3. Ns means the difference is not statistically significant. * p < 0.05.

Article Snippet: Human CSCC cell line SiHa (ATCC, USA) was used.

Techniques: Over Expression, Expressing, Transfection

Journal: Cancer Management and Research

Article Title: LncRNA SLC16A1-AS1 Suppresses Cell Proliferation in Cervical Squamous Cell Carcinoma (CSCC) Through the miR-194/SOCS2 Axis

doi: 10.2147/CMAR.S276629

Figure Lengend Snippet: Overexpression of SLC16A1-AS1 led to upregulated SOCS2 through miR-194. ( A ) The expression of SOCS2 in CSCC and paired non-tumor tissues from CSCC patients were determined by RT-qPCR. ( B and C ) The effects of overexpression of SLC16A1-AS1 and miR-194 on the expression of SOCS2, which is a target of miR-194, were analyzed by RT-qPCR ( B ) and Western blot ( C ). n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human CSCC cell line SiHa (ATCC, USA) was used.

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Western Blot

Journal: Cancer Management and Research

Article Title: LncRNA SLC16A1-AS1 Suppresses Cell Proliferation in Cervical Squamous Cell Carcinoma (CSCC) Through the miR-194/SOCS2 Axis

doi: 10.2147/CMAR.S276629

Figure Lengend Snippet: SLC16A1-AS1 suppressed SiHa cell proliferation through the miR-194/SOCS2 axis. ( A – D ) Colony formation assay was performed to evaluate the effects of si-SLC16A1-AS1, miR-194 inhibitor and si-SOCS2 on the proliferation of SiHa cells. n = 3. ( E and F ) CCK-8 assay was used to detect the effects of si-SLC16A1-AS1, miR-194 inhibitor and si-SOCS2 on the proliferation of SiHa cells. n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human CSCC cell line SiHa (ATCC, USA) was used.

Techniques: Colony Assay, CCK-8 Assay

Journal: Immune Network

Article Title: TXNIP Suppresses Lung Cancer Progression by Inhibiting TRAF6-Mediated NF-κB Activation and Autophagy

doi: 10.4110/in.2025.25.e33

Figure Lengend Snippet: (A) HEK293T cells were transfected with Mock, HA-Ub, Flag-TRAF6, or different concentrations of Myc-TXNIP, as indicated. At 24 h after transfection, transfected cells were used for protein extraction. Proteins were immunoprecipitated with an anti-Flag Ab. IB assay was then performed with anti-Flag, anti-Myc, or anti-HA Ab. (B) HEK293T cells were transfected with Mock, Flag-TRAF6, HA-Ub, different concentrations of Flag-TXNIP, or Myc-TAK1, as indicated. At 24 h after transfection, transfected cells were used for protein extraction. Proteins were then immunoprecipitated with anti-Myc Ab. IB assay was performed with anti-Myc, anti-Flag, anti-TRAF6, or anti-HA Ab. (C) 293/hTLR4A cells were transfected with Mock or different concentrations of Flag-TXNIP, along with the pBIIx-luc NF-κB-dependent reporter construct and the Renilla luciferase vector. At 24 h post transfection, cells were treated or not with LPS (500 ng/ml) for 6 h and lysed. Luciferase activity was measured using a dual luciferase assay kit. Results are presented as ± SD in three independent experiments. (D) A schematic model showing how TXNIP inhibits NF-κB activation through the TRAF6-TAK1 signaling axis. * p<0.05, ** p<0.01, and *** p<0.001.

Article Snippet: TXNIP human Tagged ORF clone (#RG210804) and TAK1 (MAP3K7) (#RG213823) human Tagged ORF clone were purchased from Origene (Rockville, MD, USA).

Techniques: Transfection, Protein Extraction, Immunoprecipitation, Construct, Luciferase, Plasmid Preparation, Activity Assay, Activation Assay

Journal: Immune Network

Article Title: TXNIP Suppresses Lung Cancer Progression by Inhibiting TRAF6-Mediated NF-κB Activation and Autophagy

doi: 10.4110/in.2025.25.e33

Figure Lengend Snippet: (A, B) Ctrl H1299 and TXNIP -KO H1299 cells were treated with vehicle (DMSO, 0.1% v/v concentration), LPS (15 µg/ml), or Poly(I:C) (20 µg/ml) for different time periods (A). The residual gap between migrating cells from the opposing wound edge is expressed as a percentage of the initial scraped area (± SD, n=3 different plates) (B). (C, D) Ctrl H1299 and TXNIP -KO H1299 cells were treated with vehicle (DMSO, 0.1% v/v concentration), LPS (15 µg/ml), or Poly(I:C) (15 µg/ml) (C). The number of migrating cells was counted. Results are presented as mean ± SD of 3 independent experiments (D). (E, F) Ctrl H1299 and TXNIP -KO H1299 cells were treated with vehicle (DMSO, 0.1% v/v concentration), LPS (15 µg/ml), or Poly(I:C) (10 µg/ml). Anchorage-independent colony formation assay was performed (E), as described in Materials and Methods. The number of colonies was measured using Adobe Photoshop software. Results are presented as mean ± SD of 3 independent experiments (F, n=3 plates). (G, H) Ctrl H1299 and TXNIP -KO H1299 cells were treated with vehicle (DMSO, 0.1% v/v concentration), LPS (5 µg/ml, G), or Poly(I:C) (4 µg/ml, H). MTT assay was then performed as described in in Material and Methods. Results are presented as mean ± SD of 3 independent experiments. (I, J) 3D spheroid formation assays were performed using Ctrl A549 and TXNIP -KO A549 cells, which were seeded in 96-well plates and incubated at 37°C for 48 h to allow spheroid formation. Spheroids were then treated with either LPS (3 µg/ml) or Poly(I:C) (1 µg/ml) and cultured further as indicated. Spheroid size was measured using ImageJ software, and images were captured using phase-contrast microscopy (I, scale bar=100 µm). Data are presented as mean ± SD (J, n=5). Statistical significance (Student’s t -test). (K) A schematic model illustrating how TXNIP inhibits lung cancer progression: Upon TLR3 or TLR4 stimulation, TXNIP binds to TRAF6, preventing its dimerization and auto-ubiquitination. This inhibition suppresses both the TRAF6-BECN1 axis for autophagy and the TRAF6-TAK1 signaling axis for NF-κB activation, ultimately reducing lung cancer progression. * p<0.05, ** p<0.01, *** p<0.001, and **** p<0.0001. # p<0.05 and ## p<0.01 for comparisons between TXNIP -KO A549 and Ctrl A549 cells (in ).

Article Snippet: TXNIP human Tagged ORF clone (#RG210804) and TAK1 (MAP3K7) (#RG213823) human Tagged ORF clone were purchased from Origene (Rockville, MD, USA).

Techniques: Concentration Assay, Colony Assay, Software, MTT Assay, Incubation, Cell Culture, Microscopy, Ubiquitin Proteomics, Inhibition, Activation Assay

Journal: Molecular Medicine Reports

Article Title: A preliminary study of the effect of ECRG4 overexpression on the proliferation and apoptosis of human laryngeal cancer cells and the underlying mechanisms

doi: 10.3892/mmr.2015.4059

Figure Lengend Snippet: Sequences of primers for reverse transcription-quantitative polymerase chain reaction.

Article Snippet: Cells grown on coverslips were fixed in 4% paraformaldehyde for 15 min, permeabilized with 0.1% Triton X-100 for 30 min and incubated with rabbit anti-human anti-ECRG4 polyclonal antibody (1:100; cat. no. sc-135139; Santa Cruz Biotechnology, Inc.) at 4°C overnight.

Techniques: Sequencing

Journal: Molecular Medicine Reports

Article Title: A preliminary study of the effect of ECRG4 overexpression on the proliferation and apoptosis of human laryngeal cancer cells and the underlying mechanisms

doi: 10.3892/mmr.2015.4059

Figure Lengend Snippet: Selection of cell lines. The expression levels of ECRG4 in Hep-2 and LSC-1 human laryngeal cancer cell lines were examined by western blot analysis. Grayscale analysis was performed using β-actin as the internal control. Experimental data are expressed as the mean ± standard deviation. * P<0.05, compared with Hep-2 cells. ECRG4, human esophageal cancer-related gene 4.

Article Snippet: Cells grown on coverslips were fixed in 4% paraformaldehyde for 15 min, permeabilized with 0.1% Triton X-100 for 30 min and incubated with rabbit anti-human anti-ECRG4 polyclonal antibody (1:100; cat. no. sc-135139; Santa Cruz Biotechnology, Inc.) at 4°C overnight.

Techniques: Selection, Expressing, Western Blot, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: A preliminary study of the effect of ECRG4 overexpression on the proliferation and apoptosis of human laryngeal cancer cells and the underlying mechanisms

doi: 10.3892/mmr.2015.4059

Figure Lengend Snippet: Establishment of a cell line stably overexpressing ECRG4. (A) Western blot analysis of ECRG4 protein expression. Grayscale analysis was conducted using β-actin as the internal control. (B) Reverse transcription-quantitative polymerase chain reaction analysis of ECRG4 mRNA expression. (C) Immunofluorescence examination of ECRG4 distribution. ECRG4 expression was visible under a fluorescence microscope (red). The nuclei were stained blue. Representative results of several replica experiments are shown in the figure (magnification, x600). Experimental data are expressed as the mean ± standard deviation. ** P<0.01, compared with the pcDNA3.1 group. ECRG4, human esophageal cancer-related gene 4.

Article Snippet: Cells grown on coverslips were fixed in 4% paraformaldehyde for 15 min, permeabilized with 0.1% Triton X-100 for 30 min and incubated with rabbit anti-human anti-ECRG4 polyclonal antibody (1:100; cat. no. sc-135139; Santa Cruz Biotechnology, Inc.) at 4°C overnight.

Techniques: Stable Transfection, Western Blot, Expressing, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Immunofluorescence, Fluorescence, Microscopy, Staining, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: A preliminary study of the effect of ECRG4 overexpression on the proliferation and apoptosis of human laryngeal cancer cells and the underlying mechanisms

doi: 10.3892/mmr.2015.4059

Figure Lengend Snippet: Overexpression of ECRG4 inhibited the proliferation of laryngeal cancer cells. (A) Examination of the proliferative capacity of laryngeal cancer cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cells from the three experimental groups were seeded into 96-well plates. Five replica wells were set up for each experimental group. The OD 490 values were measured at days 0, 1, 2, 3 and 4 after cell inoculation. (B) Determination of the clonogenic capacity of laryngeal cancer cells using colony formation assay. Cells from the experimental groups were seeded into Petri dishes. After day 14 of culture, the cells were fixed and the colony formation rates were calculated under a microscope. (C) Analysis of the cell cycle by flow cytometric analysis. Experimental data are presented as the mean ± standard deviation. * P<0.05, and ** P<0.01, compared with the pcDNA3.1 group. ECRG4, human esophageal cancer-related gene 4.

Article Snippet: Cells grown on coverslips were fixed in 4% paraformaldehyde for 15 min, permeabilized with 0.1% Triton X-100 for 30 min and incubated with rabbit anti-human anti-ECRG4 polyclonal antibody (1:100; cat. no. sc-135139; Santa Cruz Biotechnology, Inc.) at 4°C overnight.

Techniques: Over Expression, Colony Assay, Microscopy, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: A preliminary study of the effect of ECRG4 overexpression on the proliferation and apoptosis of human laryngeal cancer cells and the underlying mechanisms

doi: 10.3892/mmr.2015.4059

Figure Lengend Snippet: Overexpression of ECRG4 induced apoptosis in laryngeal cancer cells through regulation of the expression of apoptosis-related factors. (A) Examination of apoptosis by flow cytometric analysis. The figure shows the representative results of several repeated experiments. (B) Examination of apoptosis by Hoechst staining. Apoptotic cells exhibited intensive chromatin condensation and dense, strongly Hoechst stained nuclei (magnification, ×400). (C) Examination of the expression of Bax, Bcl-2, cleaved-caspase-3 and cleaved-PARP by western blot analysis. Grayscale analysis was performed using β-actin as the internal control. Experimental data are expressed as the mean ± standard deviation. Compared with the pcDNA3.1 group, ** P<0.01. ECRG4, human esophageal cancer-related gene 4; Bax, Bcl-2-associated X protein; Bcl-2, anti-B-cell lymphoma 2; PARP, poly ADP-ribose polymerase.

Article Snippet: Cells grown on coverslips were fixed in 4% paraformaldehyde for 15 min, permeabilized with 0.1% Triton X-100 for 30 min and incubated with rabbit anti-human anti-ECRG4 polyclonal antibody (1:100; cat. no. sc-135139; Santa Cruz Biotechnology, Inc.) at 4°C overnight.

Techniques: Over Expression, Expressing, Staining, Western Blot, Control, Standard Deviation

Journal: Journal of Cellular and Molecular Medicine

Article Title: EN2 Regulates Pancreatic Cancer Initiation, Progression, and Epithelial‐Mesenchymal Transition Through the Notch Signalling Pathway

doi: 10.1111/jcmm.71158

Figure Lengend Snippet: The expression of EN2 in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. (A), Protein expression of EN2 in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. Crude proteins were isolated, and EN2 expression was measured by Western blot analysis. β‐Actin was used as a loading control. (B), Expression of EN2 mRNA in HPNE, pancreatic cancer cell lines, and pancreatic CSCs. RNA was isolated, and EN2 expression was measured by q‐RT‐PCR. GAPDH was used as an internal control. Data represent mean ( n = 4) ± SD. *, # and % = significantly different from HPNE ( p < 0.05). (C), Expression of EN2. Immunocytochemistry was performed to examine EN2 expression in HPNE, PANC‐1, and AsPC‐1 cells.

Article Snippet: Human pancreatic cancer stem cells (CSCs) were obtained from Celprogen and cultured in a well‐defined medium according to the manufacturer's instructions.

Techniques: Expressing, Isolation, Western Blot, Control, Reverse Transcription Polymerase Chain Reaction, Immunocytochemistry

Journal: Biomolecules

Article Title: MicroRNAs and Stem-like Properties: The Complex Regulation Underlying Stemness Maintenance and Cancer Development

doi: 10.3390/biom11081074

Figure Lengend Snippet: MicroRNAs expressed in ESCs and CSCs and their relative functions.

Article Snippet: Normal colon stem cells (Human T4056 cells, Applied Biological Materials Inc., Richmond, BC, Canada) also express high levels of miR-137 , which is downregulated in colon CSCs (EpCAM+/CD44+/CD66a − , human SW480 cells, ATCC) [ ].

Techniques: Isolation, Modification, Over Expression, FACS, Biomarker Discovery, Expressing, Inhibition, Binding Assay, Knock-Out, Migration, Activation Assay

Journal: Biomolecules

Article Title: MicroRNAs and Stem-like Properties: The Complex Regulation Underlying Stemness Maintenance and Cancer Development

doi: 10.3390/biom11081074

Figure Lengend Snippet: Noncanonical miRNAs acting in ESCs and CSCs.

Article Snippet: Normal colon stem cells (Human T4056 cells, Applied Biological Materials Inc., Richmond, BC, Canada) also express high levels of miR-137 , which is downregulated in colon CSCs (EpCAM+/CD44+/CD66a − , human SW480 cells, ATCC) [ ].

Techniques: Derivative Assay, Expressing, In Vivo, Knockdown, Inhibition

Journal: Journal of Advanced Research

Article Title: Baicalin attenuates the immune escape of oral squamous cell carcinoma by reducing lactate accumulation in tumor microenvironment

doi: 10.1016/j.jare.2025.01.021

Figure Lengend Snippet: Baicalin repressed the proliferation and migration of OSCC cells. (A) Baicalin features a flavonoid structure with formula C21H18O11. (B) The viability of OSCC cells (SCC25, CAL27) and normal (HNOK) cells was tested by baicalin administration in the concentration range of 0–200 μmol/L. (C, D) The viability analysis of OSCC cells was performed by CCK-8 assay at different baicalin concentration (12.5 μmol/L, 25 μmol/L, 50 μ mol/L). (E) The transwell migration analysis revealed the migrative ability of OSCC cells with baicalin treatment. (F) The colony formation assay and (G) EdU proliferation assay were performed to reflect the role of baicalin on the proliferative ability of OSCC cells in a concentration-dependent manner. * p < 0.05.

Article Snippet: OSCC human tumor cells (SCC25, RRID: CVCL_1682; CAL27, RRID: CVCL_1107), mice oral cancer cells (RRID: CVCL_V412) and normal oral keratinocytes cells (HNOK) were purchased from ATCC and Fenghui Biotechnology Co., Ltd (Changsha, Hunan, China).

Techniques: Migration, Concentration Assay, CCK-8 Assay, Colony Assay, Proliferation Assay

Journal: Journal of Advanced Research

Article Title: Baicalin attenuates the immune escape of oral squamous cell carcinoma by reducing lactate accumulation in tumor microenvironment

doi: 10.1016/j.jare.2025.01.021

Figure Lengend Snippet: Baicalin targeted LDHA via m 6 A-dependent pathway in OSCC. (A) The binding mode of baicalin and LDHA was analyzed by molecular docking. (B) Baicalin-LDHA interaction was verified by using the MST technology. (C) MeRIP-Seq data was shown by IGV (Integrative Genomics Viewer), and revealed the potential m 6 A modified sites on the LDHA gene. The m 6 A motif on the LDHA gene was GGAC. (D) m 6 A modification analysis was performed in OSCC cells with the baicalin administration. (E, F) RIP-PCR assay was performed in OSCC cells (SCC25, CAL27) using anti-m 6 A antibody to verify the interaction within anti-m 6 A antibody and LDHA mRNA. (G) Online immune analysis ( https://cistrome.shinyapps.io/timer/ ) revealed the correlation of LDHA with CD8 + T cells’ infiltration. (G) Prognosis analysis showed the survival rate of OSCC patients with high/low-expression of LDHA. * p < 0.05, ** p < 0.01.

Article Snippet: OSCC human tumor cells (SCC25, RRID: CVCL_1682; CAL27, RRID: CVCL_1107), mice oral cancer cells (RRID: CVCL_V412) and normal oral keratinocytes cells (HNOK) were purchased from ATCC and Fenghui Biotechnology Co., Ltd (Changsha, Hunan, China).

Techniques: Binding Assay, Modification, Expressing