PTEN基因对卵巢癌耐药细胞系C13K顺铂耐药性的影响

目的:探讨第10号染色体上PTEN对人卵巢癌顺铂耐药细胞株C13K顺铂耐药性的逆转作用。方法:将野生型PTEN基因真核表达质粒在脂质体介导下转染人卵巢癌耐药细胞系C13K细胞,同时以转染空载体和未转染C13K细胞作为对照,分别应用逆转录聚合酶链式反应(RT-PCR)及蛋白印迹法(W estern blot)检测PTEN mRNA及蛋白表达水平,对转染细胞株进行四甲基偶氮噻唑蓝(MTT)实验,观察PTEN基因对肿瘤细胞生长的抑制作用和C13K细胞对顺铂药物敏感性的影响,流式细胞仪(FACS)分析细胞的凋亡。结果:转染48 h后,与对照组相比,转染野生型PTEN基因能明显增加C13K细胞PTEN mRNA和蛋白的表达,差异有统计学意义(P<0.05);MTT法显示,转染野生型PTEN基因的C13K细胞生长明显慢于转染空载体和未转染的C13K细胞,转染PTEN的C13K细胞对顺铂的敏感性显著增加;FACS分析显示,顺铂作用24h后,转染PTEN基因能够显著提高C13K细胞凋亡率。结论:转染野生型PTEN质粒能有效地提高C13K细胞内PTEN的表达,恢复细胞对顺铂的敏感性。

Effects of wild-type PTEN gene on resistance to cisplatin in multidrug resistant human ovarian cancer cell line C13K

Objective:To evaluate the effect of wild-type PTEN gene on reversing cisplatin resistance of multidrug resistant human ovarian cancer cell line C13K.Methods:Recombinant eukaryotic expression plasmid containing human wild-type PTEN gene was transfected into multidrug resistant human ovarian cancer cell line C13K by lipofectamine 2000.The expression of PTEN were monitored by RT-PCR and Western blot in PTEN transfected and untransfected C13K cells and the apopototic rates of both cells were detected by flowcytometry after having been treated with cisplatin.Proliferation and chemosensitivity of C13K cells to cisplatin were measured by MTT assay.Results:Compared with the control,the expression of PTEN mRNA and protein were obviously increased.The growth of wild-type PTEN-transfected C13K cells was obviously slower than those of empty-vector transfected and untransfected cells.The chemosensivity of C13K cells to cisplatin was enhanced after wild-type PTEN gene was transfected into C13K cells.The apopototic rates of wild-type PTEN transfected,empty vector transfected and non-transfected cells were(41.65±0.87)%,(18.61±0.70)% and(15.28±0.80)% respectively,the difference was statistically significant(P<0.05).Conclusion:Transfection of PTEN can increase the expression of PTEN and restore drug sensitivity to cisplatin in multidrug-resistant human ovarian cancer cell line C13K.