MiR-519d represses ovarian cancer cell proliferation and enhances cisplatin-mediated cytotoxicity in vitro by targeting XIAP.
MiR-519d represses ovarian cancer cell proliferation and enhances cisplatin-mediated cytotoxicity in vitro by targeting XIAP.
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DOI：
10.2147/ott.s60289
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发表时间：
2014
影响因子：
4
通讯作者：
Liu P
中科院分区：
文献类型：
--
作者：
Pang Y;Mao H;Shen L;Zhao Z;Liu R;Liu P
MicroRNA（miRNAs）是一类非编码小RNA，在转录后水平上作为基因表达的负调控因子，在肿瘤发生和发展中起重要作用。本研究旨在探讨miR-519d在卵巢癌中的表达及其抗肿瘤作用。通过TaqMan定量逆转录-聚合酶链反应（TaqMan qRT-PCR; Life Technologies，卡尔斯巴德，CA，USA）检测卵巢癌细胞和组织中miR-519 d的表达水平。通过3-（4，5-二甲基噻唑-2-基）-2，5-二苯基四氮唑溴化物、流式细胞术和Western印迹分析miR-519 d对卵巢癌细胞增殖和顺铂化疗敏感性的影响。采用荧光素酶报告基因检测miR-519 d在X-linked inhibitor of apoptosis protein（XIAP）3 ′端非翻译区的结合位点。采用实时荧光定量聚合酶链反应（qRT-PCR）和Western blotting检测XIAP mRNA和蛋白的表达水平。miR-519d在人卵巢癌细胞系和组织中显著下调。卵巢癌细胞中miR-519 d的过表达降低了细胞增殖，并使卵巢癌细胞对顺铂诱导的细胞死亡敏感，同时增加了caspase 3的活化和聚（腺苷二磷酸[ADP]-核糖）聚合酶1的裂解。生物信息学分析表明XIAP是miR-519d的一个推定靶点。过表达miR-519d可降低XIAP在蛋白和mRNA水平的表达。相反，抑制miR-519d增加XIAP表达。荧光素酶报告基因分析证实XIAP是miR-519d的直接靶点。卵巢癌细胞系和组织中XIAP mRNA和蛋白表达水平与miR-519 d表达呈负相关。这些发现表明，miR-519d通过靶向XIAP转录物抑制细胞增殖并使卵巢癌细胞对顺铂诱导的细胞死亡敏感，表明miR-519d在人卵巢癌中发挥肿瘤抑制作用，并突出了miR-519d在卵巢癌治疗中的治疗潜力。
MicroRNAs (miRNAs) are small, noncoding RNAs that are believed to play fundamental roles in tumorigenesis and tumor development at the posttranscriptional level, as negative regulators of gene expression. This study was designed to evaluate the expression and anticancer effect of miR-519d in ovarian cancer. The expression levels of miR-519d in ovarian cancer cells and tissues were detected by TaqMan quantitative reverse transcriptase-polymerase chain reaction (TaqMan qRT-PCR; Life Technologies, Carlsbad, CA, USA). The effects of miR-519d on ovarian cancer cell proliferation and cisplatin chemosensitivity were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide, flow cytometry, and Western blotting assay. A luciferase reporter assay was performed to validate the miR-519d binding sites on the 3′ untranslated region of X-linked inhibitor of apoptosis protein (XIAP). The expression levels of XIAP mRNA and protein were examined by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting assay, respectively. miR-519d was significantly downregulated in human ovarian cancer cell lines and tissues. Overexpression of miR-519d in ovarian cancer cells decreased cell proliferation and sensitized ovarian cancer cells to cisplatin-induced cell death accompanied by increased activation of caspase 3 and cleavage of poly(adenosine diphosphate [ADP]-ribose) polymerase 1. Bioinformatics analysis indicated that XIAP was a putative target of miR-519d. Overexpression of miR-519d decreased XIAP expression at both the protein and mRNA levels. In contrast, inhibition of miR-519d increased XIAP expression. Luciferase reporter assay confirmed XIAP as a direct target of miR-519d. XIAP mRNA and protein expression levels were inversely correlated with miR-519d expression in ovarian cancer cell lines and tissues. These findings indicate that miR-519d suppresses cell proliferation and sensitizes ovarian cancer cells to cisplatin-induced cell death by targeting the XIAP transcript, suggesting that miR-519d plays a tumor-suppressive role in human ovarian cancer and highlighting the therapeutic potential of miR-519d in ovarian cancer treatment.