【摘要】 目的:观察PTEN(phosphatase and tensin hom o logy deleted on chrom osom e 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ZR-75-1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pBP-w t-PTEN和pBP-G 129R-PTEN导入人乳腺癌ZR-75-1细胞(质粒转染成功后实验分为C-W T-PTEN组、C-G 129R-PTEN组和未转染质粒组即对照组)后,以RT-PCR、W estern b lot分析目的基因的表达,并采用M TT法和流式细胞术检测细胞增殖和细胞周期。结果:C-W T-PTEN组、C-G 129R-PTEN组细胞PTEN mRNA及PTEN蛋白出现明显的高表达。C-W T-PTEN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(P<0.05)。但C-G 129R-PTEN组细胞生长的抑制率与对照组比较,差异无显著性(P>0.05)。流式细胞术显示C-W T-PTEN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。更多还原

【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 ZR-75-1.Methods: The eukaryotic expression vector pBP-wt-PTEN and pBP-G129R-PTEN containing PTEN cDNA was transfected into ZR-75-1 cells by lipofectamine.Three groups were set: C-WT-PTEN,C-G129R-PTEN and cells without transfection as the control group.Reverse transcription-polymerase chain reaction and Western-blot methods were used to detect PTEN expression in the transfected ZR-75-1 cells.The cell cycle,cell proliferation and cell apoptosis of the ZR-75-1 cells was determined by the MTT and flow cytometry.Results: High expression of PTEN mRNA and protein was seen in the transfected ZR-75-1 cells. The proliferation of C-WT-PTEN cells were inhibited by 42.7%（vs the control group,P< 0.05）,while C-G129R-PTEN cells showed no significant change in proliferation（vs the control group,P> 0.05）.The cell cycle from G₁ to S of C-WT-PTEN cells was inhibited.Conclusion: Wide type PTEN gene might suppress the growth and induces the apoptosis in human breast cancer cell line ZR-75-1 cells through its phosphatase activity.更多还原