肿瘤坏死因子相关凋亡诱导配体抑制卵巢癌3AO细胞增殖并促进细胞凋亡  

TRAIL inhibits proliferation and promotes apoptosis of 3AO ovarian cancer cells

作　　者：张少华[1] 刘凯歌[2] 成碧萍 高倩[1] 王莉[3] 杨筱凤[4] 

机构地区：[1]西安医学院第一附属医院妇科,陕西西安710077 [2]西安医学院第一附属医院消化内科,陕西西安710077 [3]西安医学院第一附属医院科研科,陕西西安710077 [4]西安交通大学医学院第一附属医院妇产科,陕西西安710061

出　　处：《细胞与分子免疫学杂志》2014年第5期453-457,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：陕西省自然科学基础研究计划(2013JM4058);陕西省教育厅项目(11JKO724);西安医学院科研项目(10FC07);西安医学院附属医院科研项目(XYFY10-11)

摘　　要：目的研究肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导卵巢癌3AO细胞凋亡的分子机制。方法用(12.5、25.0、50.0)ng/mL的重组人TRAIL蛋白与卵巢癌3AO细胞共孵育72 h,观察细胞的形态变化,在不同时间段收集细胞,MTT法检测细胞生长增殖,计算细胞的生长抑制率,annexin V-FITC/PI染色结合流式细胞术检测细胞凋亡率和细胞周期,原位末端标记法(TUNEL)观察凋亡形态特征,Western blot法检测凋亡相关蛋白caspase-3蛋白表达。结果各浓度组的TRAIL都可以引起3AO细胞形态学改变,对细胞增殖均有抑制作用(P<0.05),25 ng/mL和50 ng/mL TRAIL处理的3AO细胞出现明显的细胞凋亡和周期阻滞,随着药物作用时间的延长,G1期细胞比例逐渐增多,而S期和G2/M期细胞比例减少,caspase-3蛋白呈高表达,但两组之间的凋亡率和凋亡蛋白的表达无显著性差异(P>0.05)。结论 TRAIL是通过抑制细胞周期、阻滞DNA合成、活化caspase-3诱导卵巢癌3AO细胞凋亡。Objective To investigate the effect of tumor necrosis factOr-related apoptosis-inducing ligand （TRAIL） on the proliferation of ovarian carcinoma 3AO cells in vitro and its molecular mechanism. Methods Ovarian carcinoma 3AO cells were incubated with 12.5, 25.0, 50.0 ng/mL TRAIL proteins for 72 hours, respectively. At 24, 48 and 72 hours, the cells were collected to observe the morphological change under an inverted microscope, detect the cell proliferation using methyl thiazolyl tetrazolium （MTT） assay, the apoptosis rate and cell cycle using flow cytometry （ FCM）, the morphological features of apoptotic cells using TdT-mediated-dUTP nick end labeling （TUNEL） and the expression of caspase-3 protein using Western blotting. Results The growth of 3AO cells was inhibited by different concentrations of TRAIL （P 〈 0. 05）. Morphological change of 3AO cells was clearly observed. TRAIL protein at 25 and 50 ng/mL significantly induced 3AO cell apoptosis and cell cycle arrest. With the treatment time went by, the percentage of cells at G1 phase increased and cells at S and G2/M phase decreased. The expression of caspase-3 protein was raised by TRAIL. No significant differences were noted in the apoptosis rate and the expression of caspase-3 protein between the 25 and 50 ng/mL TRAIL groups （P 〉 0. 05 ）. Conclusion TRAIL protein could promote the apoptosis of 3AO cells through inhibiting cell growth cycle, blocking DNA synthesis and activating caspase-3.

关 键 词：TRAIL 3AO细胞 细胞凋亡 细胞周期 CASPASE-3蛋白 

分 类 号：R392.11[医药卫生—免疫学] Q253[医药卫生—基础医学]