Department of Obstetrics and Gynecology, Renmin Hospital of Wuhan University, Wuhan 430060, P.R. China.
Mol Med Rep. 2013 Feb;7(2):425-30. doi: 10.3892/mmr.2012.1216. Epub 2012 Dec 3.
Survivin (SVV) is an important member of the inhibitor of apoptosis family. It is overexpressed in a number of cancer types, including human ovarian carcinomas. SVV promotes invasion, metastasis, growth and survival of malignant cells and confers resistance to specific chemotherapeutic drugs. The present study aimed to elucidate the role and possible mechanisms of SVV in cisplatin-resistant ovarian cancer cells (A2780/CP). Using a loss-of-function approach, we investigated the effects of adenovirus-mediated knockdown of SVV by small hairpin RNA (ad5-SVV) on the expression of pro-caspase-3, cleaved caspase-3, proliferating cell nuclear antigen (PCNA) and matrix metalloproteinase-2 (MMP-2) in A2780/CP cells by real-time PCR and western blot analysis. Proliferation was measured by MTT assay, invasive potential by Transwell, and cell apoptosis by FITC-Annexin V and propidium iodide for the functional analysis of A2780/CP cells following infection with ad5-SVV. As a result, knockdown of SVV downregulated the expression of PCNA and MMP-2 and upregulated the expression of pro-caspase-3 and cleaved caspase-3. In addition, knockdown of SVV enhanced cisplatin-induced proliferative activities, induced cell apoptosis and inhibited the invasive potential in A2780/CP cells. The present findings demonstrate that knockdown of SVV contributes to antitumor activity in cisplatin-resistant ovarian cancer cells via the downregulation of PCNA and MMP-2 expression and the upregulation of caspase-3 expression and indicate that SVV is a potential target for therapeutic anticancer drugs.
存活素 (SVV) 是凋亡抑制因子家族的重要成员。它在许多癌症类型中过度表达,包括人类卵巢癌。SVV 促进恶性细胞的侵袭、转移、生长和存活,并赋予其对特定化疗药物的耐药性。本研究旨在阐明 SVV 在顺铂耐药卵巢癌细胞 (A2780/CP) 中的作用及其可能机制。我们采用失活功能方法,通过小发夹 RNA (ad5-SVV) 腺病毒介导的 SVV 敲低,研究了 SVV 敲低对 A2780/CP 细胞中原胱天蛋白酶-3、裂解胱天蛋白酶-3、增殖细胞核抗原 (PCNA) 和基质金属蛋白酶-2 (MMP-2) 的表达的影响实时 PCR 和 Western blot 分析。通过 MTT 测定法测量增殖,通过 Transwell 测量侵袭潜力,通过 FITC-Annexin V 和碘化丙啶测量细胞凋亡,以对感染 ad5-SVV 的 A2780/CP 细胞进行功能分析。结果表明,SVV 敲低下调了 PCNA 和 MMP-2 的表达,上调了原胱天蛋白酶-3 和裂解胱天蛋白酶-3 的表达。此外,SVV 敲低增强了顺铂诱导的 A2780/CP 细胞的增殖活性,诱导细胞凋亡并抑制其侵袭潜力。这些发现表明,SVV 敲低通过下调 PCNA 和 MMP-2 的表达以及上调 caspase-3 的表达,有助于顺铂耐药卵巢癌细胞的抗肿瘤活性,并表明 SVV 是潜在的治疗性抗癌药物靶点。