辐射诱导表达载体pshuttle-Egr1-shTRAIL对肝癌SMMC7721细胞生长的抑制作用  

Inhibitory effects of recombinant plasmid pshuttle-Egr1-shTRAIL transfection in combination with X-irradiation on growth of liver cancer cells SMMC7721

作　　者：陈志勇[1,2] 刘敏[2] 龚守良[3] 董丽华[2] 

机构地区：[1]内蒙古民族大学附属医院肿瘤科,内蒙古通辽028000 [2]吉林大学第一医院肿瘤中心放疗科,吉林长春130021 [3]吉林大学公共卫生学院卫生部放射生物学重点实验室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2011年第6期981-984,共4页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(30570546)

摘　　要：目的:研究携带可溶性人肿瘤坏死因子相关凋亡诱导配体(shTRAIL)基因的辐射诱导表达载体pshuttle-Egr1-shTRAIL在人肝癌细胞株中的表达及其促凋亡作用。方法:体外通过脂质体转染SMMC7721细胞。转染细胞分别接受0(假照组)、2、5和10Gy X线照射,ELISA法检测sTRAIL的表达;细胞分为正常SMMC7721组,分别接受0(假照组)、2和5Gy X线照射的转染空质粒pshuttle-Egr1组及转染重组质粒pshutlle-Eyr1-shTRAIL组,采用AnnexinV-FITC凋亡检测试剂盒检测细胞凋亡率;分别取SMMC772细胞、转染STRAIL细胞、转染空载体及照射组细胞,采用克隆形成实验检测细胞存活率。结果:不同剂量X射线照射SMMC7721-shTRAIL细胞上清中可溶性TRAIL表达量明显高于假照组(P<0.001);与假照组比较,照射转染后SMMC7721细胞的凋亡细胞百分数明显增加(P<0.05或P<0.001),细胞存活率明显下降(P<0.05或P<0.001)。结论:pshuttle-Egr1-shTRAIL载体联合放射治疗能够显著抑制肝癌细胞SMMC7721细胞的生长,并促进其凋亡。Objective To investigate the effects of recombinant plasmid pshuttle-Egr1-shTRAIL stable transfection in combination with X-ray irradiation on the TRAIL protein expression and the apoptosis in human SMMC7721 hepatoma cells.Methods The pshuttle-Egr1-shTRAIL packaged with liposome was stably transfected into SMMC7721 cells in vitro.The shTRAIL protein expressions were measured with ELISA assay,Annexin V-FITC kit was adopted to measure the apoptosis of pshuttle-Egr1-shTRAIL cells,and the changes in survival rate of SMMC7721 cells were measured with cell cloning assay.Results The TRAIL protein expressions in pshuttle-Egr1-shTRAIL plus different doses of irradiation groups were significantly increased compared with 0 Gy group(P< 0.001).The percentage of apoptotic cells was significantly higher than that in 0 Gy group(P<0.05 or P<0.001),and the survival rate of SMMC7721 cells was decreased significantly(P<0.05 or P<0.001).Conclusion The pshuttle-Egr1-shTRAIL stable transfection in combination with irradiation can significantly induce the apoptosis of SMMC7721 tumor cells and inhibit the cell proliferation.

关 键 词：肿瘤坏死因子相关凋亡诱导配体 基因克隆 肝肿瘤 辐射 早期生长反应1 

分 类 号：R735[医药卫生—肿瘤]