生存素反义核酸诱导肝癌细胞株SMMC-7721凋亡和增加其对抗肿瘤药物敏感性的研究

Effect of Survivin-antisense on Liver Cancer Cell Line SMMC-7721 in Inducing Apoptosis and Increasing its Sensitivity to Antineoplastic Agents

背景:生存素是凋亡抑制蛋白家族的重要成员,生存素基因有可能成为肿瘤反义基因治疗的理想靶基因。目的:研究生存素反义核酸诱导肝癌细胞株SMMC-7721凋亡和增加其对常用抗肿瘤药物敏感性的作用,探讨生存素反义核酸用于肿瘤基因治疗的可能性。方法:应用基因重组技术构建pEGFP-C1-生存素反义核酸重组质粒,以脂质体转染法转染SMMC-7721细胞,用逆转录聚合酶链反应(RT-PCR)检测生存素mRNA的表达,用流式细胞仪检测细胞凋亡。将生存素反义核酸分别与7种常用抗肿瘤药物共同作用于SMMC-7721细胞,用四唑蓝(MTT)比色法测定细胞杀伤率。结果:生存素反义核酸可抑制SMMC-7721细胞中生存素mRNA的表达,从而导致细胞凋亡增加,其作用呈剂量依赖性。生存素反义核酸可增加SMMC-7721细胞对7种常用抗肿瘤药物的敏感性,明显增强药物的杀伤作用。结论:生存素反义核酸能靶向抑制野生型生存素基因的表达,提高肝癌细胞对常用抗肿瘤药物的敏感性,有可能成为肿瘤基因治疗的新方法。

Background: Survivin is an important family member of inhibitors of apoptosis proteins, and sur-vivin gene may be served as an ideal target gene in the antisense gene therapy of cancers. Aims: To study the effect of survivin-antisense on liver cancer cell line SMMC-7721 in inducing apoptosis and increasing its sensitivity to the common antineoplastic agents, and to investigate the possibility of survivin-antisense in the gene therapy of cancers. Methods: Recombinant pEGFP-Cl-survivin-antisense plasmid was obtained by gene reconstruction, and transfected to SMMC-7721 cells by lipofectamine. Expression of survivin mRNA was determined by reverse transcriptase polymerase chain reaction (RT-PCR), and apoptosis by flow cytometry in SMMC-7721 cells. Methyl thiazolyl tetrazolium (MTT) method was used to detect the killing rates of SMMC-7721 cells after treatment with survivin-antisense combined with 7 common antineoplastic agents, respectively. Results: The expression of survivin mRNA was decreased and cell apoptosis increased in SMMC-7721 cells in a dose-dependent manner after transfection of survivin-antisense. The sensitivity of SMMC-7721 cells to the 7 common antineoplastic agents and the killing rates were also increased respectively after transfection. Conclusions: Transfection of survivin-antisense can inhibit the expression of wild type survivin gene and increase the sensitivity of liver cancer cells to common antineoplastic agents. It may be used as a new gene therapy in the future treatment of cancers.