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(P<0.05);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(P<0.05)。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。
Background and purpose：miR-101 has been reported to be down-regulated in gastric cancer, colorectal cancer, breast cancer as well as prostate cancer acting as a tumor suppressor gene. However, its function in ovarian cancer is still unknown. The aim of this study was to investigate whether miR-101 can suppress cell growth and invasion of ovarian cancer cells by targeting DNMT3A, so as to reveal molecular mechanism to inhibit ovarian cancer. Methods：Quantitative real-time palymerase chain reaction （qRT-PCR） method was employed to detect the expression of miR-101 in ovarian cancer and cancer adjacent normal ovarian tissues. SKOV3 cells were transfected with miR-101 mimics, and DNMT3A siRNA was transfected as a positive control. Then Western blot was used to detect the expres-sion of DNMT3A protein regulated by miR-101 in SKOV3 cells. The growth and invasion ability of SKOV3 cells were evaluated by MTT and Transwell invasion assays.Results：qRT-PCR showed that miR-101 was down-regulated in ovarian cancer tissues. Western blot showed that the level of DNMT3A protein was inhibited by restored miR-101 or knock-down of DNMT3A in SKOV3 cells. Following transfection of miR-101 mimics or knock-down of DNMT3A for 48, 72 and 96 h respectively, MTT assay showed that theD values were signiifcantly lower than the control group, （P〈0.05）. After transfection of miR-101 mimics or knock-down of DNMT3A for 36 h, Transwell invasion assay showed that the numbers of cells through the basement membrane was （105±7） and （107±13）, respectively, which are signiifcantly different from the control group （213±11）, indicating invasion of SKOV3 cells signiifcantly slowed down （P〈0.05）.Conclusion：miR-101 suppresses cell growth and invasion by targeting DNMT3A in ovarian cancer.
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2015年第10期791-795,共5页
China Oncology
基金
湖南省教育厅课题资助项目(13C929)
