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目的 探讨野生型 p5 3基因对卵巢癌细胞生长抑制及凋亡的作用 ,为卵巢癌的基因治疗提供实验依据。方法 构建野生型 p5 3基因重组腺病毒载体、体外转染卵巢癌细胞株CaOV3;应用生化染色、免疫组化、聚合酶链技术 ,检测外源基因的转染率及表达效果 ;应用细胞计数、MTT法 (四甲基偶氮唑盐微量酶反应比色法 )、流式细胞术和TUNEL技术 (TDT -mediateddUTP -biotinendlabeling) ,检测细胞生长及凋亡的情况。 结果 野生型p5 3基因重组腺病毒载体可以有效地转染卵巢癌细胞株CaOV3,转染后的CaOV3细胞内可以检测到 p5 3基因的cDNA及p5 3蛋白的表达 ;转染后的CaOV3细胞生长受到明显抑制 ,6 3%的细胞生长停滞于G0 /G1期 ,4 0 %～ 5 0 %的细胞TUNEL呈阳性。结论 野生型 p5 3基因的导入 ,可抑制卵巢癌细胞的生长并诱导细胞凋亡。野生型 p5 3基因导入可能成为今后临床基因治疗卵巢癌的可行性方法之一。
Objective To investigate the effect of wild-type p5 3 gene on the growth inhibition and apoptosis of ovarian cancer cells and to provide experimental evidence for the gene therapy of ovarian cancer. Methods The wild-type p53 gene was constructed and transfected into ovarian cancer cell line CaOV3 in vitro. The transfection efficiency and expression of exogenous gene were detected by biochemical staining, immunohistochemistry and polymerase chain reaction. MTT assay (MTT micro-enzyme reaction colorimetric method), flow cytometry and TUNEL technology (TDT -mediated dUTP-biotinendlabeling), detection of cell growth and apoptosis. Results The wild-type p53 gene recombinant adenovirus vector was successfully transfected into ovarian cancer cell line CaOV3. The transfected CaOV3 cells could detect the cDNA and p5 3 protein expression of p5 3 gene. After transfection, the growth of CaOV3 cells Under the condition of obvious suppression, 63% of cells arrested in G0 / G1 phase and 40% ~ 50% of cells showed positive TUNEL. Conclusion The introduction of wild type p5 3 gene can inhibit the growth of ovarian cancer cells and induce apoptosis. The introduction of wild-type p5 3 gene may become one of the feasible methods for future clinical gene therapy of ovarian cancer.