PUMA/ASPP_1双抑癌基因对胃癌SGC-7901细胞增殖的影响  

Effects of PUMA/ASPP_1 Gene on Proliferation of Gastric Cancer SGC-7901 Cells

作　　者：李军[1] 龚辉[2] 朱水山[1] 

机构地区：[1]南昌大学第二附属医院消化内科,南昌330006 [2]南昌大学第四附属医院感染性疾病科,南昌330013

出　　处：《南昌大学学报（医学版）》2015年第2期5-8,61,共5页Journal of Nanchang University：Medical Sciences

基　　金：江西省教育厅科学技术研究项目(赣教技字【2006】65号)

摘　　要：目的探讨p53正向细胞凋亡调控因子(p53up-regulated modulator of apoptosis protein,PUMA)/p53促凋亡蛋白(apoptosis stimulating protein of p53,ASPP)1融合基因及单个基因对胃癌SGC-7901细胞的杀伤作用。方法通过脂质体(lipofectamine)2000TM将PUMA/ASPP1融合基因及PUMA、ASPP1单个基因分别转染胃癌SGC-7901细胞系[实验设5组,分别为胃癌细胞无转染空白对照(SGC-7901)组,空载质粒转染细胞对照(SGC-7901-pcDNA3.1)组以及重组ASPP1质粒转染细胞实验(SGC-7901-ASPP1)组、重组PUMA质粒转染细胞实验(SGC-7901-PUMA)组、重组PUMA/ASPP1质粒转染细胞实验(SGC-7901-PUMA/ASPP1)组],再次经G418筛选,获得稳定表达融合基因SGC-7901-PUMA/ASPP1细胞株。通过MTT法及使用流式细胞仪测定各组胃癌SGC-7901系细胞的存活数(光吸收度即A值)及细胞周期。结果 SGC-7901组与SGC-7901-pcDNA3.1组A值比较差异无统计学意义(P>0.05);SGC-7901-ASPP1组、SGC-7901-PUMA组和SGC-7901-PUMA/ASPP1组A值明显低于SGC-7901组与SGC-7901-pcDNA3.1组(均P<0.01);SGC-7901-ASPP1组、SGC-7901-PUMA组和SGC-7901-PUMA/ASPP13组中,SGC-7901-PUMA/ASPP1细胞组A值最低(P<0.01)。与SGC-7901组、SGC-7901-pcDNA3.1组比较,SGC-7901-ASPP1、SGC-7901-PUMA、SGC-7901-PUMA/ASPP1组的G1期明显增多,S期明显减少,尤以SGC-7901-PUMA/ASPP1组S期减少为甚(均P<0.01);SGC-7901-pcDNA3.1组与SGC-7901组比较G1期、S期差异无统计学意义(P>0.05)。结论双抑癌基因可有效抑制胃癌SGC-7901系细胞增殖,双抑癌基因较单个抑癌基因具有更强的抗肿瘤作用。ABSTRACT:Objective To investigate the lethal effects of p53 up-regulated modulator of apopto-sis(PUMA),apoptosis stimulating protein of p53(ASPP1 )and PUMA-ASPP1 fusion gene on hu-man gastric cancer SGC-7901 cells.Methods PUMA gene,ASPP1 gene and PUMA-ASPP1 fusion gene were transfected into SGC-7901 cells by using the Lipofectamine 2000 TM,respectively. SGC-7901 cells were divided into five groups:no treatment(SGC-7901 group),empty plasmid transfection(SGC-7901-pcDNA3.1 group),recombinant ASPP1 plasmid transfection(SGC-7901-ASPP1 group),recombinant PUMA plasmid transfection(SGC-7901-PUMA group),and recombi-nant PUMA-ASPP1 plasmid transfection(SGC-7901-PUMA-ASPP1 group).The stable SGC-7901-PUMA-ASPP1 cells were obtained by G418 growth inhibition screen.Cell survival and cell cycle were examined by MTT and flow cytometry,respectively.Results There were no significant differences in the A value(light absorbance)and number of cells in G1/S phase between SGC-7901 group and SGC-7901-pcDNA3.1 group (P > 0.05 ).Compared with SGC-7901 group and SGC-7901-pcDNA3.1 group,the A value reduced,number of cells in G1 phase increased and num-ber of cells in S phase decreased in SGC-7901-ASPP1 group,SGC-7901-PUMA group and SGC-7901-PUMA-ASPP1 group(P <0.01).The minimum A value and number of cells in S phase were observed in SGC-7901-PUMA-ASPP1 group.Conclusion The PUMA-ASPP1 fusion gene is more effective than PUMA or ASPP1 alone for inhibiting the proliferation of gastric cancer SGC-7901 cells.

关 键 词：P53 正向细胞凋亡调控因子 p53促凋亡蛋白 胃癌细胞 增殖 抑制 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学] R735.2[医药卫生—基础医学]