Journal: Oncology Reports

Article Title: miR-1299/NOTCH3/TUG1 feedback loop contributes to the malignant proliferation of ovarian cancer

doi: 10.3892/or.2020.7623

Figure Lengend Snippet: miR-1299 is downregulated in OC and acts as a negative regulator of NOTCH3. (A) Analysis of miR-1299 expression in OC and normal ovary tissues (NC) using RT-qPCR. (B) miR-1299 expression was significantly lower in OC tissues with high NOTCH3 expression compared to those with low NOTCH3 expression. (C) Correlation between miR-1299 and NOTCH3 levels in OC cell lines. (D) RT-qPCR analysis of NOTCH3 pathway genes after transfection of miR-1299 mimics in A2780 cells. (E) Western blot analysis of NOTCH3 after transfection of miR-1299 mimics in A2780 and CAOV3 cell lines. (F) Putative miR-1299 binding sequence in NOTCH3 3′UTR (untranslated region) (WT) and the mutated 3′UTR sequence (MUT). (G) Relative luciferase activity of reporter plasmids carrying wild-type (WT) or mutant (MUT) NOTCH3 3′UTR in A2780 cells co-transfected with NC or miR-1299 mimics. Means ± SD are shown. Statistical analysis was conducted using Student's t-test. *P<0.05 and **P<0.01; ns, not significant. OC, ovarian cancer; NOTCH3, notch receptor 3; HES1, hairy and enhancer of split-1; HEY1, Hes related family BHLH transcription factor with YRPW motif 1; HEY 2, Hes related family BHLH transcription factor with YRPW motif 2; JAG1, Jagged canonical Notch ligand 1; PBX1, Pre-B-cell leukemia transcription factor 1.

Article Snippet: The human OC cell lines A2780, CAOV3, and SKOV3 were purchased from the National Infrastructure of Cell Line Resources in China (Beijing, China).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Binding Assay, Sequencing, Luciferase, Activity Assay, Mutagenesis

Journal: Oncology Reports

Article Title: miR-1299/NOTCH3/TUG1 feedback loop contributes to the malignant proliferation of ovarian cancer

doi: 10.3892/or.2020.7623

Figure Lengend Snippet: Upregulation of miR-1299 inhibits cell proliferation, colony formation, EdU incorporation, and cell cycle in OC A2780 and CAOV3 cell lines. (A) RT-qPCR analysis of miR-1299 expression level in OC cells after transfection of NC or miR-1299 mimics for 48 h. (B) Cell proliferation of OC cells after transfection of NC or miR-1299 mimics by CCK-8 assay. (C) Representative photographs and quantifications of the colony formation assay after transfection of NC or miR-1299 mimics. (D) Representative photographs of EdU incorporation assay in OC cells after transfection of NC or miR-1299 mimics. (E) Cell cycle distribution of OC cells after transfection of NC or miR-1299 mimics by flow cytometry. Means ± SD are shown. Statistical analysis was conducted using Student's t-test. *P<0.05 and **P<0.01, miR-1299 mimic compared with the NC mimic; ns, not significant; OC, ovarian cancer.

Article Snippet: The human OC cell lines A2780, CAOV3, and SKOV3 were purchased from the National Infrastructure of Cell Line Resources in China (Beijing, China).

Techniques: Quantitative RT-PCR, Expressing, Transfection, CCK-8 Assay, Colony Assay, Flow Cytometry

Journal: Oncology Reports

Article Title: miR-1299/NOTCH3/TUG1 feedback loop contributes to the malignant proliferation of ovarian cancer

doi: 10.3892/or.2020.7623

Figure Lengend Snippet: lncRNA TUG1 functions as a sponge of miR-1299 and promotes cell proliferation in OC. (A) Screening methods for regulatory lncRNAs of miR-1299 in OC. (B) The two highest scoring putative miR-1299 binding sites in the TUG1 sequence (WT) and the point mutations in either binding site (MUT). (C) Relative luciferase activity of reporter plasmids carrying wild-type (WT) or mutant (MUT) TUG1 binding sites in A2780 cells co-transfected with NC or miR-1299 mimics. (D) Confirmation of TUG1 knockdown in A2780 cells transfected with TUG1 shRNA by RT-qPCR analysis. (E) RT-qPCR analysis of miR-1299 level in A2780 cells transfected with TUG1 shRNA or scrambled shRNA. Results of the (F) cell proliferation, (G) colony formation, (H) EdU incorporation assays, and (I) cell cycle analysis of A2780 cells transfected with NC-sh, TUG1-sh1, TUG1-sh2, and TUG1-sh+miR-1299 inhibitors. (J) Representative blot image of NOTCH3 protein level in A2780 cells transfected with NC-sh, TUG1-sh1, TUG1-sh2, and TUG1-sh+miR-1299 inhibitors by western blotting. Means ± SD are shown. Statistical analysis was conducted using Student's t-test and one-way ANOVA with Tukey post hoc test. *P<0.05 and **P<0.01; ns, not significant; OC, ovarian cancer; NOTCH3, notch receptor 3; TUG1, lncRNA taurine upregulated gene 1.

Article Snippet: The human OC cell lines A2780, CAOV3, and SKOV3 were purchased from the National Infrastructure of Cell Line Resources in China (Beijing, China).

Techniques: Binding Assay, Sequencing, Luciferase, Activity Assay, Mutagenesis, Transfection, Knockdown, shRNA, Quantitative RT-PCR, Cell Cycle Assay, Western Blot

Journal: Oncology Reports

Article Title: miR-1299/NOTCH3/TUG1 feedback loop contributes to the malignant proliferation of ovarian cancer

doi: 10.3892/or.2020.7623

Figure Lengend Snippet: TUG1 is a potential downstream target of NOTCH3. (A) Schematic diagram showing RBP Jκ motifs around the transcriptional start site (TSS) of the TUG1 gene. (B and C) TUG1 expression was inversely correlated with NOTCH3 mRNA in fresh OC tissues (B) and in the TCGA database (C). (D) RT-qPCR analysis of TUG1 level in A2780 cells treated with DAPT. (E) Diagrams showing the miR-1299/NOTCH3/TUG1 feedback loop in the development of OC. Statistical analysis was conducted using one-way ANOVA with Tukey post hoc test and Pearson's correlation analysis. **P<0.01; ns, not significant; OC, ovarian cancer; NOTCH3, notch receptor 3; TUG1, lncRNA taurine upregulated gene 1; NICD3, NOTCH3 intracellular domain; HES1, hairy and enhancer of split-1; HEY1, Hes related family BHLH transcription factor with YRPW motif 1; HEY 2, Hes related family BHLH transcription factor with YRPW motif 2.

Article Snippet: The human OC cell lines A2780, CAOV3, and SKOV3 were purchased from the National Infrastructure of Cell Line Resources in China (Beijing, China).

Techniques: Expressing, Quantitative RT-PCR

Journal: Journal of Ovarian Research

Article Title: Fisetin-induced cell death in human ovarian cancer cell lines via zbp1-mediated necroptosis

doi: 10.1186/s13048-022-00984-4

Figure Lengend Snippet: Fisetin inhibited the proliferation of both human ovary carcinoma cell lines (A2780 and OVCAR-3) in a dose-dependent manner. (A) The chemical structure of fisetin. (B) The MTT assay indicated that fisetin reduced the proliferation of both OC cell lines after treatment at incremental concentrations (25, 50 and 100 mmol/L) for 72 h. (C) Quantification of vital cells after fisetin treatment

Article Snippet: Human ovarian carcinoma (OC) cell lines were obtained from Procell (Procell, China) and cultured in DMEM (HyClone, USA) supplemented with 10% fetal bovine serum (Gibco, USA) and 1% penicillin–streptomycin (Sigma, USA).

Techniques: MTT Assay