E2F陷阱DNA对前列腺癌PC-3M细胞凋亡的诱导  

Apoptosis of the prostate cancer cell line PC-3M induced by E2F decoy DNA

作　　者：王涛[1] 姜安丽[1] 张鹏举[1] 陈彤[2] 贺美兰[1] 陈蔚文[1] 邓景惕[1] 张建业[1] 

机构地区：[1]山东大学医学院生物化学教研室 [2]济南中医院,山东济南250012

出　　处：《中国病理生理杂志》2006年第8期1462-1466,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30470820)

摘　　要：目的:为观察转录因子E2F陷阱DNA对雄激素非依赖性前列腺癌细胞PC-3M增殖和凋亡的影响。方法:采用脂质体lipofectam ine将E2F decoy DNA、ARE decoy DNA和control decoy DNA分别转染PC-3M细胞,MTT检测其对细胞增生的影响,倒置相差显微镜观察细胞形态变化,流式细胞术检测细胞凋亡率,并进行染色体DNA断裂的测定;通过RT-PCR检测转染的PC-3M细胞中c-Myc mRNA、cyc lin D1 mRNA表达水平的变化,通过W estern b lotting检测细胞中c-Myc蛋白、cyc lin D1蛋白表达水平的变化。结果:E2F decoy DNA转染后的PC-3M细胞的生长受到明显抑制;转染后的细胞形态变化符合凋亡的典型改变,染色体断裂明显,细胞凋亡率为26.35%;c-Myc、cyc lin D1的表达受到抑制。结论:E2F decoy DNA可诱导雄激素非依赖性前列腺癌PC-3M凋亡和抑制细胞增殖,其机制可能涉及到c-Myc mRNA、cyc lin D1的表达变化。AIM： To observe the effect of E2F decoy DNA on proliferation and apoptosis of androgen - independent prostate cancer cell line PC-3M. METHODS： E2F decoy DNA, ARE decoy DNA and control decoy DNA were transfected into PC -3M cells with lipofectamine, respectively. Their effects on cell proliferation were detected by MTT assay. The changes of cell morphology were observed by inverted phase contrast microscope. The cell apoptotic rate was determined by flow cytometry （FCM） analysis and chromosome DNA ladder was detected by DNA gel electrophoresis. The expression of c - Myc mRNA and cyclin DI mRNA was detected by RT - PCR. The protein levels of c - Myc and cyclin DI were detected by Western blotting. RESULTS： The growth of PC - 3M cells was inhibited after transfection. The transfected PC -3M cells displayed typical apoptotic morphological changes. The apoptotic rate was 26. 35% and DNA ladder was observed after transfection. The expression of c - Myc and cyclin DI were inhibited. CONCLUSION： These results indicate that E2F decoy DNA induces apoptosis of androgen - independent prostate cancer cell lines PC -3 M and inhibits cell proliferation via inhibiting expression of c - Myc and cyclin DI.

关 键 词：转录因子E2F 前列腺肿瘤 细胞凋亡 PC-3M细胞 

分 类 号：R363[医药卫生—病理学]