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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 member of the apoptosis-inhibiting protein family. Survivin gene may be an ideal target gene for antisense gene therapy. OBJECTIVE: To investigate the effect of survivin antisense nucleic acid on inducing hepatocellular carcinoma cell line SMMC-7721 apoptosis and increasing its sensitivity to commonly used anti-tumor drugs, and to explore the possibility of survivin antisense nucleic acid for gene therapy of cancer. Methods: Recombinant plasmid pEGFP-C1-survivin antisense was constructed by gene recombination and transfected into SMMC-7721 cells by lipofectamine. The expression of survivin mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) The apoptosis was detected by flow cytometry. Survivin antisense nucleic acids were respectively combined with 7 kinds of commonly used anti-tumor drugs in SMMC-7721 cells, and cell killing rate was determined by MTT assay. Results: Survivin antisense nucleic acid could inhibit the expression of survivin mRNA in SMMC-7721 cells, leading to the increase of apoptosis, and its effect was dose-dependent. Survivin antisense nucleic acid can increase the sensitivity of SMMC-7721 cells to 7 kinds of commonly used anti-tumor drugs, significantly enhance the killing effect of the drug. CONCLUSIONS: Survivin antisense nucleic acid can inhibit the expression of wild-type survivin gene and improve the sensitivity of hepatocarcinoma cells to commonly used anti-tumor drugs, which may become a new method for gene therapy of cancer.