CUGBP1基因对EGCG诱导人肺腺癌A549细胞增殖和凋亡作用研究  

Proliferation and apoptosis of CUGBP1 gene in human lung adenocarcinoma A549 cells induced by epigallocatechin-3-gallate

作　　者：张静[1] 曹留霞 刘晓萍[1] 姚如永[2] 荆昭[1] 张代军[1] 

机构地区：[1]青岛大学医学院组织胚胎学教研室,山东青岛266021 [2]青岛大学医学院附属医院中心实验室,山东青岛266021

出　　处：《中华肿瘤防治杂志》2014年第1期20-24,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：山东省自然科学基金(ZR2012CM008)

摘　　要：目的:研究表没食子儿茶素-3-没食子酸酯(epigallocatechin-3-gallate,EGCG)对人肺癌A549细胞增殖和凋亡影响及与CUGBP1表达的关系。方法:MTT法检测不同剂量EGCG对A549细胞增殖活性的影响;流式细胞仪检测细胞凋亡率;免疫组化和蛋白质印迹法检测各处理组CUGBP1蛋白的表达;Real-Time PCR对CUGBP1mRNA表达进行定量分析。结果:EGCG显著抑制A549细胞增殖活性,F=1 096.65,P<0.001;EGCG孵育4和24hA549细胞的凋亡率分别为(34.10±1.10)%和(51.62±1.50)%,较4和24h正常对照组的(3.90±0.97)%和(1.53±1.11)%明显增加,差异有统计学意义,F=1 254.28,P<0.001。A549细胞细胞核和细胞质内CUGBP1蛋白表达呈阳性,EGCG孵育4h可见A549细胞核CUGBP1蛋白表达呈阴性;孵育24h,部分A549细胞变圆、核固缩,CUGBP1蛋白表达又呈阳性。CUGBP1mRNA定量分析可见,EGCG明显抑制CUGBP1表达,EGCG孵育24h,A549细胞中CUGBP1mRNA相对表达量为(0.71±0.076),较4h的(0.40±0.017)高,差异有统计学意义,t=-6.782,P=0.020。结论:EGCG干扰CUGBP1基因表达抑制人肺癌细胞株A549的增殖,可能是EGCG促进A549细胞凋亡的途径之一。OBJECTIVE： To study the effects of epigallocatechin-3-gallate （EGCG） on the proliferation of human lung cancer A549 cells and the CUGBP1 expression in it. METHODS： Proliferation activities and cell apoptosis rate of Hu- man lung cancer cell line A549 induced by EGCG at different concentration were detected respectively by MTT assay and flow cytometry. The CUGBP1 protein and CUGBPlmRNA expression of A549 cells treated with EGCG were examined respectively by immunohistochemistry （IHC）, Western Blot and Real-Time PCR. RESULTS： EGCG markedly inhibited the proliferation of A549 cells（F~ 1 096.65, P＇~0. 001）and obviously promoted the apoptosis of A549 cells, the apoptosis rate of 4 h and 24 h [respectively（34.10＋_ 1.10）%, （51.62-4-1.50）%] was statistically significant compared with 4 h and 24 h normal control group [（3.90~0.97）%,（1.53_＋1.11）%] （F=I 254.28,P^0. 001）. The CUGBP1 protein expres- sion of A549 cells in cytoplasm and nucleus were positive, the CUGBP1 protein expression in nucleus of A549 cells incuba- ted with EGCG for 4 h was negative. However,most of the A549 cells treated with EGCG for 24 h became round,nuclear pyknosis and CUGBP1 protein expression in the nucleus and cytoplasm of those cells were shown positive again. Quantita- tive analysis for the CUGBPlmRNA showed that EGCG significantly inhibited the CUGBP1 expression of A549 cells,the relative expression amount of CUGBP1 mRNA with EGCG for 24 h（0.71＋0. 076） was higher than that of 4 h（0.40~ 0. 017）, there was statistically significant difference （t =- 6. 782, P = 0. 020）. CONCLUSION： EGCG can inhibits the growth and promoted the cell apoptosis of lung cancer cell line A549, which is mediated by regulating CUGBP1 gene expression.

关 键 词：肺肿瘤 表没食子儿茶素-3-没食子酸酯 人肺腺癌细胞株A549 细胞增殖 细胞凋亡 基因 CUGBPl 免疫组 织化学 

分 类 号：R734.2[医药卫生—肿瘤]