Liu Ping, Qi Xiaorong, Bian Ce, Yang Fan, Lin Xiaojuan, Zhou Shengtao, Xie Chuan, Zhao Xia, Yi Tao
Department of Gynecology and Obstetrics, Key Laboratory of Birth Defects and Related Diseases of Women and Children of The Ministry of Education, West China Second Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
State Key Laboratory of Biotherapy and Cancer Center, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Oncol Lett. 2017 Jun;13(6):4039-4046. doi: 10.3892/ol.2017.5961. Epub 2017 Mar 31.
The role of microRNA-18a (miRNA/miR-18a) as a tumor suppressor or promoter in a number of different types of cancer has been reported. However, to date, the expression and the effects of miR-18a in epithelial ovarian cancer (EOC) remain elusive. In the present study, the expression of miR-18a in patient EOC tissues and ovarian cancer cell lines was investigated using the reverse transcription-quantitative polymerase chain reaction. Luciferase assays and western blotting were performed to detect the potential direct targets of miR-18a. An A2780cp intraperitoneal mouse model, and Cell Counting Kit 8, flow cytometry and terminal deoxynucleotidyl-transferase-mediated dUTP nick end labeling assays, were used to investigate the effect of miR-18a on tumor growth and . The results indicated that the expression of miR-18a was reduced in EOC tissue and in the investigated ovarian cancer cell lines compared with non-malignant (normal) ovarian tissues and the human ovarian epithelium cell line, respectively. Overexpression of miR-18a in the A2780s and A2780cp cell lines significantly induced cell cycle arrest and apoptosis. It was demonstrated that miR-18a directly targets tumor protein p53-regulating inhibitor of apoptosis gene 1 and inositol phosphate multikinase, hence regulating the expression of downstream targets. The A2780cp intraperitoneal mouse model was employed and the results indicated that miR-18a may inhibit A2780cp intraperitoneal tumor growth by inhibiting proliferation and inducing apoptosis. Together, the results of the present study demonstrated that miR-18a has a role as a tumor suppressor by inhibiting proliferation and inducing apoptosis. Assessment of miR-18a expression may provide a novel method for diagnosis and be a therapeutic target for EOC.
据报道,微小RNA-18a(miRNA/miR-18a)在多种不同类型癌症中具有肿瘤抑制或促进作用。然而,迄今为止,miR-18a在上皮性卵巢癌(EOC)中的表达及作用仍不明确。在本研究中,采用逆转录-定量聚合酶链反应检测患者EOC组织和卵巢癌细胞系中miR-18a的表达。进行荧光素酶报告基因检测和蛋白质免疫印迹法以检测miR-18a的潜在直接靶点。使用A2780cp腹腔内小鼠模型、细胞计数试剂盒8、流式细胞术和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记检测法,研究miR-18a对肿瘤生长及 的影响。结果表明,与非恶性(正常)卵巢组织和人卵巢上皮细胞系相比,EOC组织和所研究的卵巢癌细胞系中miR-18a的表达分别降低。在A2780s和A2780cp细胞系中过表达miR-18a可显著诱导细胞周期阻滞和凋亡。结果表明,miR-18a直接靶向肿瘤蛋白p53调节的凋亡抑制基因1和肌醇磷酸多激酶,从而调节下游靶点的表达。采用A2780cp腹腔内小鼠模型,结果表明miR-18a可能通过抑制增殖和诱导凋亡来抑制A2780cp腹腔内肿瘤生长。总之,本研究结果表明,miR-18a通过抑制增殖和诱导凋亡发挥肿瘤抑制作用。评估miR-18a的表达可为EOC的诊断提供一种新方法,并成为其治疗靶点。