PDCD4对DAP5表达及大肠癌细胞凋亡的影响  

Effects of Programmed Cell Death 4 Gene on Expression of Death Associated Protein 5 and Apoptosis in Colon Cancer Cells

作　　者：易波[1,2] 李其云[2] 饶华民[2] 邵江华[3] 

机构地区：[1]江西省分子医学重点实验室,330006 [2]江西省肿瘤医院腹部外一科 [3]南昌大学第二附属医院普外科

出　　处：《天津医药》2013年第6期520-522,共3页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(项目编号:81160309);江西省科技厅科技支撑项目-社会发展支撑计划项目(项目编号:2010BSA13900);江西省卫生厅一般科技项目(项目编号:20091215)

摘　　要：目的探讨程序性死亡基因4(PDCD4)对死亡相关蛋白5(DAP5)的表达及对人大肠癌细胞系LOVO凋亡的影响。方法构建重组真核表达载体pFLAG/PDCD4,转染人大肠癌细胞LOVO,G418(500mg/L)筛选获得稳定表达PDCD4的细胞系。RT-PCR及Western blotting检测LOVO细胞中PDCD4的mRNA及蛋白表达,流式细胞仪检测LOVO细胞凋亡情况,Western blotting检测LOVO细胞DAP5表达的变化。结果成功建立稳定表达PDCD4的大肠癌细胞LOVO-pFLAG/PDCD4。转染PDCD4的LOVO-pFLAG/PDCD4组与空白对照LOVO组、转染空载质粒的LOVO-pFLAG组相比,PDCD4蛋白表达和mRNA水平明显升高(P<0.01);细胞凋亡率明显增加(P<0.01);同时伴有DAP5蛋白表达明显升高(P<0.01)。结论 PDCD4能够诱导大肠癌细胞LOVO的凋亡,其机制可能与上调DAP5的表达有关。Objective To investigate the effect of exogenous programmed cell death 4 （PDCD4） gene on the expression of death associated protein 5 （DAPS） and apoptosis of human colorectal cancer cell LOVO, and involved mechanisms thereof. Methods Recombinant eukaryotic plasmid pFLAG/PDCD4 was constructed and transfeeted into human colorectal cancer cell LOVO. Cells stably expressing PDCD4 were established by G418 selection （500 mg/L）. The levels of PDCD4 protein and mRNA were analyzed by RT-PCR and Western blotting. The apoptotic cells were measured by flow eytometry, and protein expression of DAP5 was detected by Western blotting. Results LOVO-pFLAG/PDCD4 cell line was successfully established by G418 selection. Compared to non-transfection and mock-transfection group, the levels of PDCD4 mRNA and protein were significantly increased, the cell apoptosis ratio was enhanced and the expression of DAP5 protein was increased in transfection group （P 〈 0.01）. Conclusion PDCD4 could induce apoptosis of human colorectal cancer LOVO cells, which mechanism might be involved in up-regulating DAP5 protein.

关 键 词：蛋白激酶类 结直肠肿瘤 癌 细胞凋亡 质粒 转染 重组 遗传 基因 肿瘤抑制 程序性死亡基因4 死亡相关蛋白5 

分 类 号：R735.340.2[医药卫生—肿瘤]