摘要
目的:通过转染PTEN基因表达载体,提高宫颈癌Hela细胞中PTEN的表达水平,观察能否影响Hela细胞对顺铂的敏感性,并探讨其分子学机制。方法:将培养的宫颈癌Hela细胞分为空白对照(未转染质粒的Hela细胞)、质粒对照(转染空质粒的Hela细胞)和PTEN转染(转染PTEN表达载体的Hela细胞)3组。分别用RT-PCR和Western检测各组Hela细胞中PTEN的mRNA和蛋白的表达水平;MTT法检测各组Hela细胞对顺铂的化学敏感性;RT-PCR检测各组Hela细胞中磷脂酰肌醇3-激酶(PI3K)的表达水平。结果:PTEN转染组Hela细胞中PTEN的表达水平比另2组细胞显著增高(P<0.01);经顺铂处理后,PTEN转染组Hela细胞生长抑制率比其它2组显著增高(P<0.01),PTEN转染组Hela细胞中PI3K被显著抑制(P<0.01)。结论:PTEN能增强宫颈癌Hela细胞对顺铂的化疗敏感性,它可能通过下调Hela细胞中PI3K而导致Hela细胞对顺铂耐药性下降。
Objective:To investigate whether the chemosensitivity of cisplatin on cervical cancer Hela cell could be influenced by enhanced PTEN(phosphatase and tensin homology deletion on chromosome ten)expression from transfected PTEN vector.Methods:Three groups of Hela cell were cultured:blank control(non-transfection),plasmid control(Hela cells transfected with blank plasmid)and PTEN transfected cells(Hela cells transfected with plasmid contains PTEN gene).RT-PCR and Western blotting were used to detect and assess the expression level of the PTEN mRNA and protein in three groups of Hela cells respectively.Three groups of Hela cells were treated with different concentrations of cisplatin.MTT assay was used to assess chemosensitivity in three groups of Hela cells treated with cisplatin.RT-PCR was used to detect the expression level of phosphatidylinositol 3-kinase(PI3K)in three groups of Hela cell.Results:PTEN expression in PTEN transfected Hela cells was significantly higher than in other two groups(P0.01).After treated with cisplatin,the growth inhibition ratio of PTEN transfected Hela cells was higher than other groups of cells(P0.01).PI3K expression was significantly inhibited in PTEN transfected cells than the other two groups(P0.01).Conclusion:PTEN could increase chemosensitivity of cisplatin on cervical cancer Hela cells,which could be induced by reducing drug resistance from inhibition of PI3K in Hela cells by PTEN.
出处
《中日友好医院学报》
2012年第1期25-28,38,F0004,共6页
Journal of China-Japan Friendship Hospital