miR-101靶向甲基化转移酶3A抑制人卵巢癌细胞生长与侵袭

发布时间：2019-02-28 20:10

【摘要】：背景与目的:mi R-101在胃癌、结肠癌、乳腺癌以及前列腺癌中表达下调,有类似抑癌基因样作用,然而,其在卵巢癌中的作用尚未明确。该研究旨在探讨mi R-101是否通过靶向调控甲基化转移酶3A(DNMT3A)抑制人卵巢癌细胞生长与侵袭,从而进一步揭示mi R-101的抑瘤机制。方法:采用实时定量聚合酶链式反应(quantitative real-time polymerase chain reaction,q RT-PCR)检测22例卵巢癌组织及癌旁正常卵巢组织中mi R-101的表达改变;将mi R-101 mimics转染于卵巢癌SKOV3细胞,以DNMT3A si RNA为阳性对照,采用蛋白[质]印迹法(Western blot)检测外源过表达mi R-101对DNMT3A蛋白表达水平的影响;采用噻唑蓝(thiazolyl blue,MTT)和Transwell侵袭实验检测外源高表达mi R-101对人卵巢癌细胞生长与侵袭能力的影响。结果:q RT-PCR检测结果显示,mi R-101在22例卵巢癌组织中的表达水平较癌旁正常组织明显下调;Western blot检测结果显示,外源过表达mi R-101或沉默DNMT3A能下调SKOV3细胞DNMT3A蛋白的表达水平;MTT检测结果显示,转染mi R-101 mimics或沉默DNMT3A 48、72和96 h后D值与对照组比较明显减少,差异均有统计学意义(P0.05);Transwell侵袭实验显示,转染mi R-101 mimics或沉默DNMT3A 36 h后穿过基底膜的细胞数分别为(105±7)个和(107±13)个,与对照组(213±11)个比较能明显减缓SKOV3细胞的穿膜能力,差异有统计学意义(P0.05)。结论:mi R-101通过靶向调控DNMT3A抑制人卵巢癌细胞生长与侵袭。
[Abstract]:Background & AIM: the down-regulated expression of mi rn 101 in gastric cancer, colon cancer, breast cancer and prostate cancer has a similar anti-oncogene effect. However, its role in ovarian cancer is still unclear. The aim of this study was to investigate whether mi RX101 could inhibit the growth and invasion of human ovarian cancer cells by targeting methyltransferase 3A (DNMT3A), thus further revealing the antitumor mechanism of mi RX101. Methods: real-time quantitative polymerase chain reaction (quantitative real-time polymerase chain reaction,q RT-PCR) was used to detect the expression of mi rq101 in 22 cases of ovarian cancer and normal ovarian tissues adjacent to cancer. Human ovarian cancer cell line SKOV3 was transfected with mi rm 101 mimics, and DNMT3A si RNA was used as positive control. (Western blot) was used to detect the effect of exogenous overexpression of mi Rm 101 on the expression of DNMT3A protein. The growth and invasion ability of human ovarian cancer cells were detected by thiazolyl blue,MTT and Transwell invasion assay, and the effects of exogenous high expression of mi Rx101 on the growth and invasion ability of human ovarian cancer cells were detected. Results: the results of Q-RT-PCR showed that the expression level of mi-ru-101 in 22 cases of ovarian carcinoma was significantly lower than that of normal tissue adjacent to the tumor. The results of Western blot analysis showed that the expression level of DNMT3A protein in SKOV3 cells could be down-regulated by exogenous overexpression of mi-Rx101 or silencing DNMT3A. The results of MTT detection showed that the D value was significantly decreased after 72 h and 96 h after transfection with mi Rc 101 mimics or silencing DNMT3A 48 compared with the control group, the difference was statistically significant (P0.05). Transwell invasion assay showed that the number of cells passing through the basement membrane was (105 卤7) and (107 卤13), respectively, after transfection with mi Rc 101 mimics or silencing DNMT3A for 36 h, which was significantly lower than that of the control group (213 卤11), and the number of cells passing through the basement membrane was significantly lower than that of the control group (213 卤11). The difference was statistically significant (P0.05). Conclusion: mi Rc 101 inhibits the growth and invasion of human ovarian cancer cells by targeting DNMT3A.
【作者单位】：郴州市第一人民医院妇产科;
【基金】：湖南省教育厅课题资助项目(13C929)
【分类号】：R737.31

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