miR-129-5p靶向HMGB1对卵巢癌细胞增殖、侵袭和迁移的影响及分子机制

为探讨miR-129-5p对卵巢癌(ovarian cancer,OC)细胞增殖、侵袭和迁移的影响及分子机制,qRT-PCR检测不同OC细胞株和正常人卵巢上皮细胞株中miR-129-5p的表达水平;在SKOV3细胞中分别转染miR-129-5p模拟物或无关序列,qRT-PCR检测转染后细胞中miR-129-5p的表达情况。生物信息学软件TargetScan预测miR-129-5p与高迁移率族蛋白B1(high mobility group box 1 protein, HMGB1)基因靶向结合位点,qRT-PCR、Western blotting和双荧光素酶报告基因实验分析miR-129-5p对HMGB1的靶向调控作用。分别采用MTT和Transwell实验检测SKOV3细胞增殖、侵袭和迁移能力。将miR-129-5p模拟物和HMGB1 siRNA或siRNA Control共转染至SKOV3细胞,MTT和Transwell实验检测其对细胞增殖、侵袭和迁移能力的影响。结果显示,SKOV3细胞中miR-129-5p的表达量显著低于正常人卵巢上皮细胞(P<0.05)。在SKOV3细胞中转染miR-129-5p模拟物可显著提高miR-129-5p的表达量(P<0.05)。miR-129-5p与HMGB1 3’非翻译区存在靶向结合位点,且过表达miR-129-5p可抑制HMGB1的表达,miR-129-5p模拟物转染后相对荧光素酶活性显著下降(P<0.05)。过表达miR-129-5p后,SKOV3细胞增殖能力明显减弱,侵袭和迁移能力明显受到抑制(P<0.05)。而转染HMGB1 siRNA后,miR-129-5p模拟物对SKOV3细胞增殖、侵袭和迁移的抑制作用明显减弱(P<0.05)。以上结果表明miR-129-5p通过靶向调控HMGB1的表达抑制SKOV3细胞增殖、侵袭和迁移。

MiR-129-5p inhibits proliferation,invasion and migration of ovarian cancer cells by targeting HMGB1 and its molecular mechanism

We sought to investigate the effect of miR-129-5 p on the proliferation, invasion and migration of ovarian cancer(OC) cells and its molecular mechanism, real-time quantitative PCR(qRT-PCR) was used to detect miR-129-5 p expression in different OC cell lines and human normal ovarian epithelial cell lines;miR-129-5 p mimics or unrelated sequences were transfected into SKOV3 cells, respectively. qRT-PCR was used to measure the miR-129-5 p level in transfected cells. Bioinformatics software TargetScan predicted the targeted binding sites of miR-129-5 p and HMGB1, and qRT-PCR, Western blotting, and dual luciferase reporter gene experiments were used to analyze the targeted regulation of miR-129-5 p on HMGB1. MTT and Transwell experiments were used to detect cell proliferation, invasion, and migration ability, respectively. The miR-129-5 p mimic and HMGB1 siRNA or siRNA Control were co-transfected into SKOV3 cells, and the effects of cell proliferation, invasion, and migration ability were examined using MTT and Transwell experiments. The results showed that the expression of miR-129-5 p in SKOV3 cells was significantly lower than that of human normal ovarian epithelial cells(P < 0.05). Transfection of miR-129-5 p mimics, significantly increased the expression of miR-129-5 p in SKOV3 cells(P < 0.05). miR-129-5 p and HMGB1 3’non-codingregion have targeted binding sites, and overexpression of miR-129-5 p inhibited the expression of HMGB1, and the relative luciferase activity after transfection of miR-129-5 p mimics is significantly down-regulated(P < 0.05). After miR-129-5 p was over-expressed, the proliferation ability of SKOV3 cells was significantly weakened, and the invasion and migration ability were significantly inhibited(P < 0.05). Furthermore, transfection with HMGB1 siRNA also significantly inhibited the proliferation, invasion and migration of SKOV3 cells(P < 0.05). The above results indicate that miR-129-5 p inhibits the proliferation, invasion and migration of SKOV3 cells through targeted regulation of HMGB1 expression.