可溶性人TRAIL诱导胃癌细胞BGC-823凋亡的研究  

Soluble human TRAIL induces apoptosis of BGC-823 cells

作　　者：李彦[1] 姜政[1] 向廷秀[2] 曾波[1] 陶小红[1] 王丕龙[1] 

机构地区：[1]重庆医科大学附属第一医院消化内科 [2]重庆医科大学附属第一医院实验研究中心,重庆400016

出　　处：《第三军医大学学报》2008年第11期1066-1070,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金(30500234)~~

摘　　要：目的通过构建pBud-EGFP-TRAIL质粒体外观察可溶性人TRAIL蛋白(sTRAIL)对胃癌细胞BGC-823的作用。方法以EFGP基因为报告基因,sTRAIL为目的基因,构建pBud-EGFP-TRAIL双表达质粒,鉴定酶切和测序重组体。经脂质体转染法转染体外培养的胃癌细胞BGC-823,用荧光显微镜观察EGFP的表达,RT-PCR及Western blot进一步检测sTRAIL mRNA及其蛋白的表达,MTT法检测转染后对胃癌细胞的生长抑制率,TUNEL法观察细胞的凋亡情况,流式细胞仪分析转染后胃癌细胞的凋亡率。结果测序结果证实pBud-EGFP-TRAIL载体构建成功,质粒经脂质体转染后,通过表达sTRAIL蛋白对胃癌细胞发挥作用。MTT法显示转染该质粒的细胞的生长抑制率明显高于对照组,TUNEL法显示细胞核固缩,核染色质聚集或断裂,形成凋亡小体。流式细胞仪结果表明转染该质粒的细胞的的凋亡率明显高于对照组。结论在体外初步证实TRAIL能诱导胃癌细胞BGC-823凋亡。Objective To study the effect of soluble TRAIL protein（sTRAIL） on the gastric cancer cell line BGC-823. Methods EGFP as report gene and sTRAIL as the target gene, a double expression plasmid of pBud-EGFP-TRAIL was constructed. After identified by restriction digestion and sequencing, pBud-EGFP-TRAIL was transfected into gastric cancer cell line BGC-823 by Lipofectamine ^TM2000. The expression of EGFP was observed by fluorescent microscope. The mRNA and protein expressions of sTRAIL were detected by RT-PCR and Western blot. The proliferation of BGC-823 cells was examined by MTT method, and the apoptosis of BGC-823 cells was examined by TUNEL method and flow cytometry. Results Sequencing result showed that the recombinant plasmid of pBud-EGFP-TRAIL had been constructed successfully. After transfected with pBud- EGFP-TRAIL, BGC-823 cells expressed sTRAIL protein and their apoptosis was induced. The inhibition rate of proliferation of BGC-823 cells transfected with pBud-EGFP-TRAIL was higher than that of the control by MTT, and the apoptosis rate was also higher by flow cytometry. Karyopycnosis, nuclear chromosome condensation and nuclear segmentation were observed by TUNEL. Conclusion It is proved that sTRAIL can cause apoptosis of gastric cancer cell line BGC-823.

关 键 词：胃肿瘤 肿瘤坏死因子 肿瘤坏死因子相关诱导凋亡配体 细胞凋亡 BGC-823细胞 

分 类 号：R394.3[医药卫生—医学遗传学] R730.23[医药卫生—基础医学]