Huang Jiansheng, Lu Dudan, Xiang Tianxin, Wu Xiaoping, Ge Shanfei, Wang Yue, Wang Jiaxin, Cheng Na
Department of Infectious Diseases, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.
Oncol Lett. 2020 Apr;19(4):3173-3180. doi: 10.3892/ol.2020.11431. Epub 2020 Mar 3.
The present study investigated whether microRNA (miR)-132-3p targeted transcription factor SOX-4 (Sox4) for the inhibition of proliferation, migration, invasion and promotion of apoptosis in liver cancer (LC) cells. The expression of miR132-3p and Sox4 mRNA was evaluated by quantitative PCR and protein expression was determined by western blot analysis. Cell proliferation, apoptosis, migration, and invasion were assessed at different time points by the MTT assay, flow cytometry analysis, wound healing assay and Transwell migration assay, respectively. Bioinformatics prediction and luciferase assays were performed to validate and confirm Sox4as a potential target of miR-132p. There was a reduced expression of miR-132-3p in HepG2 and Huh7 cell lines compared with HccLM3 cells. Overexpression of miR-132-3p resulted in significant inhibition of proliferation and induction of apoptosis in LC cells. Moreover, migration and invasion of HepG2 cells were suppressed by over expressing miR-132-3p. However, downregulation of miR-132-3p in Hep-G2 cells promoted cell growth, invasion and migration and inhibited apoptosis. Bioinformatics analysis predicted Sox4 as a potential target of miR-132-3p, which was further confirmed by the luciferase reporter assay. In addition, an inverse association was observed between miR-132-3p and Sox4 expression. miR-132-3p may regulate the proliferation, apoptosis, migration and invasion of HepG2 cells by targeting Sox4.
本研究调查了微小RNA(miR)-132-3p是否靶向转录因子SOX-4(Sox4),以抑制肝癌(LC)细胞的增殖、迁移、侵袭并促进其凋亡。通过定量PCR评估miR132-3p和Sox4 mRNA的表达,并通过蛋白质印迹分析确定蛋白质表达。分别在不同时间点通过MTT法、流式细胞术分析、伤口愈合试验和Transwell迁移试验评估细胞增殖、凋亡、迁移和侵袭。进行生物信息学预测和荧光素酶测定以验证并确认Sox4是miR-132p的潜在靶点。与HccLM3细胞相比,HepG2和Huh7细胞系中miR-132-3p的表达降低。miR-132-3p的过表达导致LC细胞的增殖受到显著抑制并诱导其凋亡。此外,过表达miR-132-3p可抑制HepG2细胞的迁移和侵袭。然而,Hep-G2细胞中miR-132-3p的下调促进了细胞生长、侵袭和迁移并抑制了凋亡。生物信息学分析预测Sox4是miR-132-3p的潜在靶点,荧光素酶报告基因测定进一步证实了这一点。此外,观察到miR-132-3p与Sox4表达之间呈负相关。miR-132-3p可能通过靶向Sox4来调节HepG2细胞的增殖、凋亡、迁移和侵袭。