外源性P27kip1基因转染诱导前列腺癌细胞凋亡的实验研究

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

Experimental Study on the Induction of Prostate Cancer Apoptosis by Exogenous p27kip1 Gene Transfer

Objective To explore the novel strategy for gene therapy of prostate cancer. Methods The p27kip1 gene was transferred into prostate cancer cell line PC-3M under the induction of liposome. Exogenous p27kip1 gene expression was detected by SABC immunohistochemistry. The in vitro cellular growth activities were assayed by MTT colorimetry; cell cycle changes were detected by flow cytometry (FCM); apoptosis was assayed by DNA badder and acridine orange-ethidium bromide fluorescent staining methods. Results After exogenous p27kip1 being transferred, the p27kip1 protein levels in prostate cancer cells were obviously elevated (P<0.01), with in vitro growth being inhibited by 12.24 % to 36.52 % (P<0.01). The cell cycle was arrested at G 0/G 1 phase, with apoptosis characteristic "sub-G1" peaks. There were obvious trapeziform bands on electrophoresis, with the apoptosis rate being 18.5 % (P<0.01). Conclusion Apoptosis of prostate cancer could be significantly induced through enhancing the negative regulation on cell cycle by exogenous p27kip1 gene transfer, which was a potential method for gene therapy of prostate cancer.