摘要
目的 研究野生型PTEN体外瞬时转染对人乳腺癌MCF 7细胞的生长抑制作用和对阿霉素敏感性的增效作用。方法 构建带有人全长PTENcDNA的真核表达载体pEGFP C1 PTEN ,以脂质体介导的方法转染MCF 7细胞,观察对细胞生长的抑制作用。以MTT法检测细胞对阿霉素的敏感性。结果 PTEN使MCF 7细胞出现皱缩变圆,胞质出芽等凋亡形态学改变;流式细胞仪检测显示转染PTEN 4 8hG0 /G1期细胞增加了14 79% ,并出现凋亡峰(10 6 0 % ) ;PTEN显著降低阿霉素处理后的克隆存活率;PTEN转染组细胞对阿霉素的敏感性显著增加(χ2 =8 5 9,P <0 0 5 ) ,其半数致死剂量(IC50 )仅为空载体对照的1/ 2 (t=4 77,P <0 0 1)。结论 PTEN过表达诱导MCF
Objective To study whether transient overexpression of tumor suppressor gene PTEN could lead to growth suppression and up-regulate the sensitivity to doxorubicin of human breast cancer MCF-7 cells. Methods The eukaryotic expression plasmid pEGFP-C 1-PTEN containing whole cDNA of PTEN was constructed and transfected into MCF-7 cells by Lipofectamine 2000 in vitro. Growth inhibition of the cells was observed by phase contrast microscope and flow cytometry. The clonogenic cell survival ability was studied by clony forming assay. MCF-7 cells′ chemosensitivity to adriamycin was studied with MTT assay. Results PTEN overexpression led to morphological changes characteristic of apoptosis of MCF-7 cells. PTEN overexpression also resulted in a significant increase in G 0/G 1 cell population (14.79%) and apoptosis (10.60%) detected by flow cytometry. The clonogenic survival rate of cells transfected with PTEN was significantly decreased after doxorubicin treatment compared with control. The transfected cells were more sensitive to doxorubicin compared with the control cells ( χ 2=8.59 , P <0.05), and the IC 50 decreased by 1/2 of that of the vector control ( t=4.77, P <0.01). Conclusion Overexpression of PTEN induces apoptosis and G 1 arrest and also increases the chemosensitivity to doxorubicin in human breast cancer MCF-7 cells.
出处
《中华普通外科杂志》
CSCD
北大核心
2005年第4期240-242,共3页
Chinese Journal of General Surgery
基金
国家自然科学基金资助项目 (3 0 3 0 0 12 4)
