摘要
背景与目的:近来的研究表明PTEN基因(phosphatase and tensin homology deleted on chromosome ten,第10号染色体上磷酸酶和张力蛋白同源缺失的基因)失活在肿瘤化疗耐药发挥了重要作用。本文研究外源性PTEN基因稳定转染人卵巢癌A2780细胞后对紫杉醇敏感性的影响。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A2780细胞,同时以转染空载体和未转染细胞作为对照(3组细胞分别命名为WT-PTEN/A2780,GFP/A2780和A2780),分别应用RT-PCR和蛋白印迹技术检测各组细胞PTEN mRNA转录和蛋白表达的变化;四甲基偶氮唑蓝实验观察转染PTEN基因对肿瘤细胞生长的抑制作用和A2780细胞对紫杉醇药物敏感性的影响,流式细胞仪(FACS)分析细胞凋亡情况。结果:与对照组相比,转染野生型PTEN基因能明显增加A2780细胞PTENmRNA转录和蛋白的表达,差异有显著性(P<0.05);MTT法显示,WT-PTEN/A2780细胞生长明显慢于GFP/A2780和A2780细胞;紫杉醇对WT-PTEN/A2780,GFP/A2780和A2780的IC50值分别为(7.6±0.40)nmol/l、(22.5±0.9)nmol/l和(22.7±0.8)nmol/l,WT-PTEN/A2780细胞对紫杉醇药物敏感性显著增加(P<0.05);FACS分析显示,紫杉醇作用24h后,WT-PTEN/A2780,GFP/A2780和A2780的凋亡率分别为(34.6±1.6)%、(13.5±0.9)%和(12.7±1.0)%,差异有显著性(P<0.05)。结论:转染野生型PTEN重组质粒能有效提高A2780细胞内PTEN的表达,诱导A2780细胞凋亡,显著增加A2780细胞对紫杉醇杀伤的敏感性。
Background and purpose：Loss of PTEN （phosphatase and tensin homology deleted on.chromosome ten） plays an important role in the drug resistance of cancer.This study was designed to evaluate the effect of exogenous wild PTEN gene stable transfection on increasing sensitivity of human ovarian cancer cell line A2780 to paclitaxel. Methods：Wild-type PTEN recombinant eukaryotic expression plasmid was constructed and then was transfected into A2780 cells by lipofectamine 2000. The expression of PTEN mRNA and protein were monitored by RT-PCR and Western blot. Proliferation and chemosensitivity of cells to paclitaxel were measured by methyl thiazolyl tetrazolium（MTT）,and cell apoptosis was detected by flow cytometry after having been treated with paclitaxel. Empty plasmid-transfected A2780 and normal A2780 cells were used as control （the different there groups were named as WT-PTEN/A2780, GFP/A2780 and A2780）.Results：RT-PCR and Western blot showed that A2780 cells express high level of PTEN mRNA and protein after infection with WT-PTEN plasmid; The proliferation rate of WT-PTEN/A2780 cells was obviously slower than those of other groups; the chemosensitivity of A2780 cells to paclitaxel was enhanced after wild-type PTEN gene was transfected into A2780 cells. The apoptotic rates of WT-PTEN/A2780, GFP/A2780 and A2780 cells were （34.55±1.62）%、（13.50±0.94）% and（12.72±0.97）% respectively, the difference was statistically significant （P〈0.05）.Conclusions：Transfection of PTEN could increase the expression of PTEN and increase drug sensitivity to human ovarian cancer cell line A2780 by inducing apoptosis.
出处
《中国癌症杂志》
CAS
CSCD
2007年第9期669-673,共5页
China Oncology
基金
国家自然科学基金资助项目(30571950)
国家973重点基础研究发展计划资助项目(2002CB513107)
