外源性p27kip1基因转染诱导前列腺癌细胞凋亡的实验研究

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

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 。

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.