Overexpression of miRNA-221 promotes cell proliferation by targeting the apoptotic protease activating factor-1 and indicates a poor prognosis in ovarian cancer.
Overexpression of miRNA-221 promotes cell proliferation by targeting the apoptotic protease activating factor-1 and indicates a poor prognosis in ovarian cancer.
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DOI：
10.3892/ijo.2017.3898
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发表时间：
2017-04
影响因子：
5.2
通讯作者：
You Z
中科院分区：
文献类型：
--
作者：
Li J;Li Q;Huang H;Li Y;Li L;Hou W;You Z
MicroRNAs是一类小的非编码的内源性RNA，参与癌症的发生和发展。据报道，microRNA-221(mir-221)在人类肿瘤中具有致癌和抑瘤作用，但miR-221在卵巢癌中的作用尚不清楚。本研究采用定量逆转录聚合酶链式反应(qRT-PCR)和免疫印迹方法分别检测了63例卵巢癌组织和细胞株IOSE25、A2780、OVCAR3、SKOV3和3AO中miR-221和APAF1蛋白的表达水平。用细胞计数试剂盒-8(CCK-8)检测细胞增殖，用Transwell法检测细胞迁移和侵袭，用流式细胞仪和Hoechst染色检测细胞凋亡，用荧光素酶分析验证APAF1基因3‘-UTRmiR-221可能的靶点。MiR-221在卵巢癌组织中表达上调。与miR-221低表达患者相比，miR-221高表达患者的无病生存期(P=0.0014)和总生存期(P=0.0058)均降低。将miR-221抑制剂导入SKOV3和A2780细胞，可诱导APAF1蛋白表达，抑制细胞增殖和迁移，促进肿瘤细胞凋亡。综上所述，APAF1基因被证实是miR-221的直接靶点，过表达APAF1在体外可抑制卵巢癌细胞的增殖并诱导细胞凋亡。这些发现表明miR-221-APAF1作为一种潜在的卵巢癌新的诊断或预后生物标志物值得进一步研究。
MicroRNAs are a class of small non-coding, endogenous RNAs involved in cancer development and progression. MicroRNA-221 (mir-221) has been reported to have both an oncogenic and tumor-suppressive role in human tumors, but the role of miR-221 in ovarian cancer is poorly understood. In the present study, the expression levels of miR-221 and the apoptosis protease activating factor 1 (APAF1) protein in 63 samples of ovarian cancer tissues and the cell lines, IOSE25, A2780, OVCAR3, SKOV3 and 3AO were detected by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and western blot analysis, respectively. Cell proliferation was measured using Cell Counting kit-8 (CCK-8); cell migration and invasion were detected using a Transwell assay; cell apoptosis was evaluated by flow cytometry and Hoechst staining, and a luciferase assay was performed to verify a putative target site of miR-221 in the 3′-UTR of APAF1 mRNA. Expression of miR-221 was upregulated in ovarian cancer tissues. Patients with increased miR-221 expression levels had a reduced disease-free survival (P=0.0014) and overall survival (P=0.0058) compared with those with low miR-221 expression. Transfection of SKOV3 and A2780 cell lines with miR-221 inhibitor induced APAF1 protein expression, suppressed cell proliferation and migration and promoted tumor cell apoptosis. In conclusion, the APAF1 gene was confirmed as a direct target of miR-221 and overexpression of APAF1 suppressed ovarian cancer cell proliferation and induced cell apoptosis in vitro. These findings indicate that miR-221-APAF1 should be studied further as a potential new diagnostic or prognostic biomarker for ovarian cancer.