TTP对小鼠乳腺癌细胞系4T1凋亡的调控

摘要

目的 观察TTP(trisetetraprolin)对小鼠乳腺癌细胞系4T1凋亡的影响.方法 构建mTTP真核表达质粒PCR3.1/mTTP;脂质体转染小鼠乳腺癌4T1细胞系并筛选稳定转染克隆;用终浓度0.1μmol的细胞凋亡诱导剂星形孢菌素处理6 h,流式细胞仪检测细胞凋亡.结果 经G418筛选,成功得到稳定转染mTTP的细胞株PCR3.1+mTTP和PCR3.1(阴性对照),实时定量PCR检测mTTP的表达,4T1细胞转染PCR3.1/mTTP之后,mTTP的表达明显增高.流式细胞仪检测发现,4T1、4T1 PCR3.1和4T1 PCR3.1+mTTP 3种细胞的凋亡率差异无统计学意义,细胞凋亡诱导剂作用后PCR3.1+mTTP细胞凋亡率明显增加,凋亡率为29.5%,而4T1、4T1+PCR3.1细胞的凋亡率分别为17.0%和18.0%.结论 成功构建了TTP高表达的小鼠乳腺癌细胞系PCR3.1+mTTP,该细胞对凋亡诱导药物星形孢菌素的敏感性增加.%Objective To establish a mouse breast cancer cell line 4T1 with high TTP (trisetetraprolin)expression and to observe the effect of TTP on breast cancer cell line 4T1 apoptosis.Methods The breast cancer 4T1 cell line was stably transfected with the plasmids of PCR3.1 and PCR3.1/mTTP by Lipofectamine 2000.The cell apoptosis was detected using flow cytometry technology.Results Higher mTTP expression was detected in 4T1 cells that has been transfected with PCR3.1+mTTP plasmid using real time quantitative PCR.The apoptosis rate among 4T1,4T1 PCR3.1 and 4T1 PCR3.1+mTTP was not significant difference.But after treatment the cells with Staurosporine,a apoptosis promoting medicine,for 6 hours,apoptosis rate of 4T1 PCR3.1+mTTP cell line was increased.The apoptosis rate of 4T1,4T1 PCR3.1 and 4T1 PCR3.1+mTTP was 17%,18% and 29.5%,respectively.Conclusion mTTP could not enhance 4T1 cell apoptosis directly,but it could increase the sensitivity of 4T1 to apoptosis inducing medicine.