E6-AP对前列腺癌细胞株LNCaP细胞增殖和细胞周期的影响

Effects of E6-AP on proliferation and cell cycle of human prostate cancer LNCaP cells

目的:研究E6相关蛋白(E6-AP)对前列腺癌细胞增殖和细胞周期的影响,阐明E6-AP在雄激素受体通路中的作用机制及在前列腺癌发生发展中的作用。方法:采用电穿孔方法制备tTA及E6-AP稳定转染细胞株;利用强力霉素(DOX)的调节作用,通过MTT实验观察E6-AP过表达及低表达对LNCaP细胞增殖的影响;利用流式细胞仪进行PI单染色检测E6-AP过表达及低表达对LNCaP细胞周期进程的影响。结果:成功构建双质粒稳定转染细胞株,与E6-AP低表达的LNCaP细胞和DOX关闭表达后的稳定株比较,E6-AP过表达的LNCaP细胞数目明显增多(P<0.01),且处于S期的细胞比例明显增大(P<0.01)。结论:E6-AP过表达可促进LNCaP细胞增殖并促进细胞进入S期,提示E6-AP与雄激素受体信号通路有密切关联。

Objective To investigate the effects of E6-AP on cell proliferation and cell cycle of human prostate cancer cells and elucidate the role of E6-AP participating in androgen receptor signal pathway and in the progress of prostate cancer.Methods tTA and E6-AP overexpression LNCaP cell line was established by electroporation transfection.The cell viability was analyzed by MTT assay and cell cycle distribution was detected by flow cytometry after propidium iodide staining using the stable cell line under the regulation of Dox.Results LNCaP cell line overexpressing E6-AP was established successfully.Compared with untreated LNCaP cell and Dox treated E6-AP stable cell line,the total number and the proportion in S phase of Dox treated E6-AP stable cells were increased significantly（P0.01）.Conclusion Overexpression of E6-AP induces proliferation of LNCaP cells and increase the number of cells in S phase,indicating that E6-AP is involved in androgen receptor signal pathway.