Role of lncRNA-ATB in ovarian cancer and its mechanisms of action

Abstract

This study aimed to elucidate the role of long non-coding RNA activated by transforming growth factor-β (lncRNA-ATB) in ovarian cancer and its underlying mechanisms of action. Expression levels of lncRNA-ATB in ovarian cancer cell line SKOV3 and in a healthy human ovarian cell line were compared using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The results indicated that lncRNA-ATB was expressed at significantly higher levels in SKOV3 cells compared with the healthy cell line. After downregulation of lncRNA-ATB expression in SKOV3 cells using lncRNA-ATB-short hairpin RNA, cell proliferation, apoptosis, invasion and migration were assessed using Cell counting kit-8, Live Dead staining, Transwell assay and wound healing assay, respectively. RT-qPCR and western blotting were used to quantify the expression of signal transducer and activator of transcription 3 (STAT3), phosphorylated (p)-STAT3, and the additional epithelial to mesenchymal transition (EMT)-related proteins E-cadherin and vimentin in SKOV3 cells. LncRNA-ATB downregulation significantly reduced SKOV3 cell proliferation, invasion and migration, promoted apoptosis, decreased the expression of p-STAT3 and vimentin, and increased E-cadherin expression. Taken together, these results suggest that lncRNA-ATB downregulation can inhibit ovarian cancer cell proliferation, invasion and migration, and promote cell apoptosis. Lnc-RNA-ATB may therefore be a new target for ovarian cancer treatment.

Keywords: apoptosis; metastasis; ncRNA-ATB; ovarian cancer; proliferation.

Figure 1.

Expression of lncRNA-ATB in ovarian cancer cell line SKOV3. (A) RT-qPCR was used to detect the levels of lncRNA-ATB in SKOV3 cells and HOSEpiCs. (B) LncRNA-ATB-NC or lncRNA-ATB-shRNA was transfected into SKOV3 cells for 48 h, and the expression of lncRNA-ATB in SKOV3 cells was detected by RT-qPCR. Data were presented as the mean ± SD. Each experiment was repeated three times. **P<0.01 vs. control group. HOSEpiC, human ovarian surface epithelial cell; lncRNA-ATB, long non-coding RNA-activated by transforming growth factor-β; lncRNA-ATB-NC, long non-coding RNA-activated by transforming growth factor-β-negative control; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; SKOV3, ovarian cancer cell line.

Figure 2.

Effect of lncRNA-ATB downregulation on SKOV3 cell proliferation and apoptosis. LncRNA-ATB-NC or lncRNA-ATB-shRNA was transfected into SKOV3 cells for 48 h. (A) Cell counting kit-8 assay was used to detect cell proliferation ability. (B) Flow cytometry was used to detect the apoptosis level. Data are presented as the mean ± SD. Each experiment was repeated three times. **P<0.01 vs. control group. LncRNA-ATB, long non-coding RNA-activated by transforming growth factor-β; lncRNA-ATB-NC, long non-coding RNA-activated by transforming growth factor-β-negative control; SKOV3, ovarian cancer cell line; PI, propidium iodide.

Figure 3.

Effects of lncRNA-ATB downregulation on SKOV3 cell invasion and migration. (A) LncRNA-ATB-NC or lncRNA-ATB-shRNA was transfected into SKOV3 cells for 48 h, then scratch wound healing assay was used to detect cell migration ability. (B) Transwell assay was used to detect cell invasion ability (magnification, ×200). Data are presented as the mean ± SD. **P<0.01 vs. control group. lncRNA-ATB, long non-coding RNA-activated by transforming growth factor-β; lncRNA-ATB-NC, long non-coding RNA-activated by transforming growth factor-β-negative control; SKOV3, ovarian cancer cell line.

Figure 4.

Effect of lncRNA-ATB downregulation on the expression of epithelial to mesenchymal transition-related proteins and the STAT3 pathway activity in SKOV3 cells. (A) LncRNA-ATB-NC or lncRNA-ATB-shRNA was transfected into SKOV3 cells for 48 h, then western blotting was used to detect the protein level of p-STAT3, STAT3, E-cadherin and vimentin in SKOV3 cells. (B) Western blotting results were quantified. RT-qPCR was used to detect the mRNA expression of (C) STAT3, (D) E-cadherin and (E) vimentin in SKOV3 cells. Data are presented as the mean ± SD. Each experiment was repeated three times. *P<0.05 and **P<0.01 vs. control group. E-cadherin, epithelial cadherin; lncRNA-ATB, long non-coding RNA-activated by transforming growth factor-β; lncRNA-ATB-NC, long non-coding RNA-activated by transforming growth factor-β-negative control; SKOV3, ovarian cancer cell line; STAT-3, signal transducer and activator of transcription 3; p-STAT-3, phosphorylated signal transducer and activator of transcription 3.