Department of Obstetrics and Gynecology, Fuzhou General Hospital of Nanjing Military Command, Fuzhou, PR China.
Int J Gynecol Cancer. 2013 Sep;23(7):1191-7. doi: 10.1097/IGC.0b013e31829ea779.
Abnormal expression of miR-199a-3p, which has similar effects to oncogenes or tumor suppressor genes, can occur in various malignant tumors and is closely linked with tumor cell proliferation, invasion, and metastasis. However, its expression and effects in endometrial endometrioid adenocarcinoma (EEC) are still unclear. This study was designed to identify the impact of miR-199a-3p on the proliferation of EEC cells and its role in the carcinogenesis of EEC.
The expression levels of miR-199a-3p in EEC and paired adjacent nontumor tissues were analyzed by real-time polymerase chain reaction. The effects of miR-199a-3p on proliferation, cell cycle and apoptosis in EEC cells were analyzed in Ishikawa cells transfected with miR-199a-3p mimics and inhibitors. The target genes of miR-199a-3p were predicted using bioinformatics methods. The extent of regulation of the predicted target genes by miR-199a-3p was determined using luciferase reporter assays, Western blotting, and quantitative polymerase chain reaction. The EEC cells were pretreated with target gene-specific inhibitors to further identify the relationship between the effects of miR-199a-3p and the predicted target genes.
Compared with the adjacent tissues and normal endometrium, reduced expression of miR-199a-3p was found in human EEC specimens. Compared with the control group transfected with control microRNA mimics, the proliferative capacity of EEC cells transfected with miR-199a-3p mimics was inhibited, whereas cells transfected with miR-199a-3p inhibitors showed increased proliferation. The inhibitory effect was associated with increased cell populations at the G1-phase, and decreased cell populations at the S-phase. The results demonstrated that miR-199a-3p could inhibit the protein expression of mammalian target of rapamycin (mTOR) by targeted binding to the mTOR-3' untranslated region. Inhibition of EEC cell proliferation by miR-199a-3p was mediated by its targeted regulation of mTOR.
MiR-199a-3p inhibits tumor cell proliferation through negative regulation of mTOR expression. Restoration of intracellular miR-199a-3p levels may serve as a potential option for EEC treatment.
miR-199a-3p 的异常表达具有类似癌基因或抑癌基因的作用,可发生于多种恶性肿瘤中,与肿瘤细胞的增殖、侵袭和转移密切相关。然而,其在子宫内膜样腺癌(EEC)中的表达和作用尚不清楚。本研究旨在探讨 miR-199a-3p 对 EEC 细胞增殖的影响及其在 EEC 发生发展中的作用。
采用实时聚合酶链反应分析 EEC 及配对相邻非肿瘤组织中 miR-199a-3p 的表达水平。用 miR-199a-3p 模拟物和抑制剂转染 Ishikawa 细胞,分析 miR-199a-3p 对 EEC 细胞增殖、细胞周期和凋亡的影响。采用生物信息学方法预测 miR-199a-3p 的靶基因。通过荧光素酶报告基因检测、Western blot 和定量聚合酶链反应确定 miR-199a-3p 对预测靶基因的调控程度。用靶基因特异性抑制剂预处理 EEC 细胞,进一步鉴定 miR-199a-3p 作用与预测靶基因之间的关系。
与相邻组织和正常子宫内膜相比,人 EEC 标本中 miR-199a-3p 的表达降低。与转染对照 microRNA 模拟物的对照组相比,转染 miR-199a-3p 模拟物的 EEC 细胞的增殖能力受到抑制,而转染 miR-199a-3p 抑制剂的细胞增殖增加。这种抑制作用与 G1 期细胞群体增加和 S 期细胞群体减少有关。结果表明,miR-199a-3p 通过靶向结合 mTOR 3'非翻译区抑制哺乳动物雷帕霉素靶蛋白(mTOR)的蛋白表达。miR-199a-3p 通过靶向调控 mTOR 抑制 EEC 细胞增殖。
miR-199a-3p 通过负调控 mTOR 表达抑制肿瘤细胞增殖。恢复细胞内 miR-199a-3p 水平可能成为 EEC 治疗的一种潜在选择。