转染抑癌基因pten对人乳腺癌zr751细胞的细胞周期和增殖能力的影响精选.doc

转染抑癌基因PTEN对人乳腺癌ZR751细胞的细胞周期和增殖能力的影响 作者：李祥勇，林观平，周克元 【关键词】 人乳腺癌ZR 【摘要】目的：观察PTEN（phosphatase and tensin homology deleted on chromosome ten，第10号染色体上磷酸酶和张力蛋白同源缺失的基因）对人乳腺癌细胞ZR751细胞增殖和细胞周期的影响。方法：利用脂质体介导法将携带有野生型和突变型PTEN cDNA的真核表达载体pBPwtPTEN和pBPG129RPTEN导入人乳腺癌ZR751细胞（质粒转染成功后实验分为CWTPTEN组、CG129RPTEN组和未转染质粒组即对照组）后，以RTPCR、Western blot 分析目的基因的表达，并采用MTT法和流式细胞术检测细胞增殖和细胞周期。 结果：CWTPTEN组、CG129RPTEN组细胞PTEN mRNA 及PTEN蛋白出现明显的高表达。CWTPTEN组细胞生长的抑制率可高达42.7%，与对照组比较，差异有显著性(P0.05)。流式细胞术显示CWTPTEN组细胞周期从G1期到S期已发生抑制。结论：野生型PTEN可依赖其磷酸酶活性抑制肿瘤细胞的增殖，并最终诱导细胞凋亡。 【关键词】 人乳腺癌ZR751细胞； PTEN基因；细胞周期 【Abstract】 Objective: To investigate the effect of the PTEN (phosphatase and tensin homologue deleted on chromosome ten) gene on cell cycle and proliferation in human breast cancer cell line ZR751. Methods: The eukaryotic expression vector pBPwtPTEN and pBPG129RPTEN containing PTEN cDNA was transfected into ZR751 cells by lipofectamine. Three groups were set: CWTPTEN, CG129RPTEN and cells without transfection as the control group. Reverse transcriptionpolymerase chain reaction and Westernblot methods were used to detect PTEN expression in the transfected ZR751 cells. The cell cycle, cell proliferation and cell apoptosis of the ZR751 cells was determined by the MTT and flow cytometry. Results: High expression of PTEN mRNA and protein was seen in the transfected ZR751 cells. The proliferation of CWTPTEN cells were inhibited by 42.7% (vs the control group, P 0.05). The cell cycle from G1 to S of CWTPTEN cells was inhibited. Conclusion: Wide type PTEN gene might suppress the growth and induces the apoptosis in human breast cancer cell line ZR751 cells through its phosphatase activity. 【Key words】 ZR751; PTEN; cell cycle; proliferation PTEN（phosphatase and tensin homology deleted on chromosome ten，第10号染色体上磷酸酶和张力蛋白同源缺失的基因）是继p53基因之后新近发现的与各种肿瘤密切相关的一个抑癌基因，其突变与缺失见于多种恶性肿瘤，如前列腺癌、胶质瘤、乳腺癌等。目前发现PTEN是唯一一个编码具有双重磷酸酶活性的抑癌基因［1］，因其凭借磷酸酶活性负调控PI3K/Akt信号传导途径而备受关注。虽有研究表明，PTEN的缺失与突变与乳腺癌的发生发展密切相关［2］，然而目前有关PTEN基因对乳腺癌细胞生长抑制