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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 7细胞凋亡和周期停滞并显著增加对阿霉素杀伤的敏感性
Objective To investigate the inhibitory effect of wild-type PTEN transiently transfected on human breast cancer cell line MCF-7 and its synergistic effect on doxorubicin sensitivity. Methods The eukaryotic expression vector pEGFP C1 PTEN with human full-length PTEN cDNA was constructed and transfected into MCF-7 cells by liposome-mediated method to observe the inhibitory effect on cell growth. MTT assay cells to doxorubicin sensitivity. Results PTEN resulted in shrinkage and rounding of MCF 7 cells and morphological changes of apoptosis such as cytoplasm sprouting. Flow cytometry showed that the number of apoptotic cells increased by 14 79% in PTEN 48 hG0 / PTEN significantly reduced the survival rate of clones after adriamycin treatment. The sensitivity of cells to PTEN transfection was significantly increased (χ2 = 8 5 9, P <0 05). The median lethal dose (IC50) was only 1/2 of the empty vector control (t = 4 77, P <0 0 1). Conclusion PTEN overexpression induces MCF 7 cell apoptosis and cell cycle arrest and significantly increases the sensitivity to doxorubicin killing