人肝癌细胞肿瘤抑制基因转染对阿霉素敏感性的影响

目的为了解肿瘤抑制基因ｐ５３和Ｒｂ基因与肝癌对药物敏感性之间的关系，本实验用逆转录病毒介导将野生型ｐ５３或Ｒｂ基因导入人肝癌细胞株ＳＭＭＣ７７２１并观察其药物敏感性的改变。方法将野生型ｐ５３ｃＤＮＡ或ＲｂｃＤＮＡ克隆在逆转录病毒载体ｐＬＸＳＮ，转染人肝癌细胞株ＳＭＭＣ７７２１，筛选阳性克隆，同时以空载体ｐＬＸＳＮ和反义ｐ５３基因作为对照，免疫组化检测ｐ５３或Ｒｂ基因的表达。加入化疗药物４８ｈ后掺入３Ｈ－ＴｄＲ，１８ｈ后检测ＣＰＭ值。结果成功地构建了分别含野生型ｐ５３、Ｒｂ或反义ｐ５３的逆转录病毒载体，转染ＳＭＭＣ７７２１细胞系后获得了ｐ５３或Ｒｂ的表达；与亲代７７２１细胞相比，ｐ５３或Ｒｂ的表达阳性的７７２１细胞的生长速度均明显减慢，ｐ５３表达阳性的７７２１细胞对阿霉素的敏感性明显增强，同样条件下，Ｒｂ在７７２１细胞系内的表达则对阿霉素敏感性无影响。结论野生型ｐ５３基因转染能提高人肝癌７７２１细胞对阿霉素的敏感性

Effects of tumor suppressor gene transfection on adriamycin sensitivity for human hepatoma

Objective p53 gene not only regulate the growth of cell but also increase the chemosensitivity of tumor cell by inducing apoptosis of tumor cell.Whether Rb gene participated in the process of apoptosis or not is uncertain.Wild type p53(wt p53)and Rb gene were transduced into human hepatoma cell line SMMC7721 separately in order to determine the effect of wt p53 and Rb gene transfer on chemosensitivity of human hepatoma.Methods We constructed and packaged recombinant retroviral vector expressing human wtp53 and Rb.The recombinant virus was transduced into human SMMC7721 cell line,and the expression of p53 and Rb was detected.Then 3H was incorporated into p53 or Rb positive SMMC7721 after the treatment of adriamycin.Results Expression of both wt p53 and Rb were demonstrated by immunohistochemistry;The growth rate of both p53 and Rb transfected 7721 reduced;Wt p53 positive 7721 was more sensitive to adramycin than control cell lines,in contrast,Rb expression did not increase the chemosensitivity.Conclusion The expression of wtp53 gene can increase chemosensitivity of human hepatoma.