Department of Gynecology, University of Würzburg, D-97080 Würzburg, Germany.
Oncol Rep. 2012 Dec;28(6):2023-8. doi: 10.3892/or.2012.2041. Epub 2012 Sep 18.
Platinum resistance is the most crucial problem for treatment of ovarian cancer. Increasing evidence points towards AKT overexpression as a mechanistic reason for this clinical condition. The present study evaluates the effect of overexpression and downregulation of AKT on the sensitivity to cisplatin in a platinum-resistant human ovarian cancer cell line and the corresponding platinum-sensitive parental cell line. A2780 and A2780cis ovarian cancer cell lines were stably transfected with an AKT-sense and AKT-antisense plasmid. Successful transfection was evaluated by western blot analysis. Cytotoxic effects of cisplatin were evaluated by metabolic (MTT) and clonogenicity assays as well as by FACS analysis. AKT overexpression (confirmed by western blotting) converted platinum-sensitive A2780 into platinum-resistant cells as shown by MTT assay. Importantly, platinum resistance of A2780cis cells could be reversed by downregulation of AKT, as demonstrated by MTT and clonogenicity assays and FACS analysis. Our data provide strong evidence that cisplatin resistance in ovarian cancer is mediated by AKT overexpression and can be overcome by AKT downregulation, thus, providing a rationale for clinical phase II/III studies combining AKT inhibitors with cisplatin.
铂类耐药是卵巢癌治疗中最关键的问题。越来越多的证据表明 AKT 过表达是导致这种临床状况的机制原因。本研究评估了 AKT 过表达和下调对顺铂在铂耐药人卵巢癌细胞系和相应的铂敏感亲本细胞系中的敏感性的影响。通过 AKT-正义和 AKT-反义质粒稳定转染 A2780 和 A2780cis 卵巢癌细胞系。通过 Western blot 分析评估成功转染。通过代谢（MTT）和集落形成测定以及 FACS 分析评估顺铂的细胞毒性作用。AKT 过表达（通过 Western blot 证实）将铂敏感的 A2780 转化为 MTT 测定显示的铂耐药细胞。重要的是，AKT 下调可逆转 A2780cis 细胞的铂耐药性，如 MTT 和集落形成测定以及 FACS 分析所示。我们的数据提供了强有力的证据，表明卵巢癌中的顺铂耐药性是由 AKT 过表达介导的，可以通过 AKT 下调克服，从而为将 AKT 抑制剂与顺铂联合用于临床 II/III 期研究提供了依据。
