野生型p53基因重组质粒的构建及对人卵巢癌细胞的抑制作用

目的:研究p53基因转染对卵巢癌细胞系SKOV-3生物学行为的影响.方法:用分子克隆技术构建人野生型p53基因与真核细胞表达载体PCNDA3的重组体(PCNDA-p53),并用脂质体介导的转染技术,将其导入不表达p53的人卵巢癌SKOV-3细胞中,经筛选、鉴定后,观察细胞生长速度、集落形成能力和增殖周期的变化.结果:成功地构建了人野生型p53基因与真核细胞表达载体的重组体,转染后获得稳定表达p53的转染克隆.外源性p53基因的表达明显抑制了卵巢癌细胞的生长速度及集落形成能力,使细胞阻滞于G1期.结论:外源性野生型p53基因能抑制卵巢癌细胞的增殖.

Construction of wild-type p53 gene recombinant expression vector and its suppressive effects on human ovarian cancer cells

Objective: To study the effects of wild type p53 gene on biologic behavior in human ovarian cancer cell line SKOV 3. Methods: A human wild type p53 gene recombinant eukaryotic expression vector was constructed by cloning techniques and was introduced by lipofectin mediated gene transfection into SKOV 3 cell line which does not express endogenous p53. Then positive clones were observed for their biologic behavior after their p53 mRNA and protein expression detected by Northern blot and Western blot. Results:The wild type p53 gene recombinant eukaryotic expression vector was constructed successfully and the clones expressing p53 stablely were obtained. The speed of cell growth and ability of colony formation were suppressed by exogenous p53 gene,leading to stop growing of ovarian cancer cells at phase G 1.Conclusions:Exogenous wild type p53 gene can suppress the growth of the human ovarian cancer cells. This gene therapy based on restoration of the defective p53 function plays an important role in ovarian cancer treatment.