Department of Hepatobiliary Surgery, the Third Affiliated Hospital of Soochow University, Changzhou, Jiangsu 213003, PR China.
Oncol Rep. 2013 Mar;29(3):1183-8. doi: 10.3892/or.2012.2196. Epub 2012 Dec 18.
Survivin has become an attractive anticancer therapeutic target due to its important role in tumor cell viability and its selective expression in tumor cells. In the present study, we constructed a recombinant siRNA plasmid vector against survivin and stably transfected it into HepG2 and SMMC‑7721 hepatocellular carcinoma cells in vitro. Semi-quantitative RT-PCR and western blotting were used to determine the expression of survivin mRNA and protein, respectively. Tumor cell proliferation was assessed by trypan blue exclusion. We evaluated the change in caspase-3 activity, and the rate of cell apoptosis and the cell cycle distribution were analyzed by flow cytometry. Assessment of chemosensitivity was carried out by MTT assay. The results showed that transfection of survivin siRNA caused a significant inhibition of survivin mRNA and protein expression which was associated with cell growth inhibition, specific G0/G1 phase arrest, increased caspase-3 activity and enhanced chemosensitivity to cisplatin in both HCC cell lines. We suggest that suppression of survivin expression by RNAi attenuates the malignant phenotype of hepatocellular carcinoma cells, and may provide a novel approach for anticancer gene therapy.
Survivin 因其在肿瘤细胞存活中的重要作用及其在肿瘤细胞中的选择性表达，已成为一种有吸引力的抗癌治疗靶点。在本研究中，我们构建了针对 survivin 的重组 siRNA 质粒载体，并将其稳定转染到 HepG2 和 SMMC-7721 肝癌细胞中。半定量 RT-PCR 和 Western blot 分别用于检测 survivin mRNA 和蛋白的表达。台盼蓝排斥试验评估肿瘤细胞增殖。我们评估了 caspase-3 活性的变化，并通过流式细胞术分析细胞凋亡率和细胞周期分布。MTT 法评估化疗敏感性。结果表明，转染 survivin siRNA 可显著抑制 survivin mRNA 和蛋白表达，与细胞生长抑制、特定 G0/G1 期阻滞、caspase-3 活性增加和增强顺铂化疗敏感性相关。我们认为，RNAi 抑制 survivin 表达可减弱肝癌细胞的恶性表型，可能为抗癌基因治疗提供新途径。
