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目的 探索前列腺癌基因治疗的新途径。方法 用脂质体介导 p2 7kip1基因转染前列腺癌 PC- 3 M细胞 ,SABC免疫组化法检测癌细胞外源性 p2 7kip1基因表达 ;MTT比色分析检测癌细胞体外生长活性 ;流式细胞仪 ( FCM)分析细胞周期改变 ;DNA L adder、吖啶橙 -溴化乙啶荧光染色法检测细胞凋亡。结果 发现外源性 p2 7kip1基因转移后 ,前列腺癌细胞 p2 7kip1蛋白水平显著增强 ( P<0 .0 1) ,体外生长抑制 12 .2 4%～ 3 6.5 2 % ( P<0 .0 1) ,出现 G0 / G1 期细胞周期阻滞及凋亡性“亚 G1期”峰 ,电泳可见典型的“梯状”条带 ,凋亡比率为 18.5 % ( P<0 .0 1)。结论 转染外源性p2 7kip1基因能增强对细胞周期的负性调节 ,显著诱导前列腺癌细胞凋亡 ,是前列腺癌基因治疗的潜在途径
Objective To explore a new approach to gene therapy of prostate cancer. Methods PC3 M cells were transfected with p2 7kip1 gene by lipofectamine. The expression of exogenous p2 7kip1 gene was detected by SABC immunohistochemical method. The growth of cancer cells was detected by MTT colorimetric assay. Flow cytometry Cell cycle analysis was performed by FCM. Apoptosis was detected by DNA L adder and acridine orange-ethidium bromide staining. The results showed that the expression of p2 7kip1 protein in prostate cancer cells was significantly increased after exogenous p2 7kip1 gene transfer (P <0.01), and the in vitro growth inhibition was 12.24% -3.52% (P <0.01) (P <0.01). The cell cycle arrest at G0 / G1 phase and apoptotic “sub G1 phase” peak appeared. A typical “ladder” band was observed by electrophoresis. The apoptotic rate was 18.5% (P <0.01). Conclusion Transfection of exogenous p2 7kip1 gene can enhance the negative regulation of cell cycle and induce the apoptosis of prostate cancer cells significantly, which is a potential pathway for gene therapy of prostate cancer