抑制人高迁移率族蛋白1基因表达对胰腺癌细胞凋亡影响的研究  

Effects of HMGB1 inhibition on the apoptosis of human pancreatic cancer cells

作　　者：黄庆先[1] 贾秀梅[1] 王国斌[2] 

机构地区：[1]烟台毓璜顶医院肝胆外科,山东烟台264000 [2]武汉华中科技大学同济医学院附属协和医院普外科,湖北武汉430072

出　　处：《中华肿瘤防治杂志》2008年第15期1147-1150,共4页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:探讨反义人高迁移率族蛋白1(HMGB1)基因转染对人胰腺癌细胞的凋亡诱导作用及对Bcl-2/baxmRNA表达水平的影响。方法:应用分子克隆技术构建反义HMGB1基因真核表达载体pcDNA3.1/anti-HMGB1,脂质体介导转染人胰腺癌细胞panc1,G418筛选获得阳性克隆。RT-PCR方法检测HMGB1、Bcl-2和Bax表达水平,MTT法测定细胞增殖,流式细胞仪(FCM)检测细胞凋亡及细胞周期情况。结果:成功构建pcDNA3.1/anti-HMGB1真核表达载体,转染pcDNA3.1/anti-HMGB1可使panc1细胞HMGB1和Bcl-2mRNA表达水平显著降低,P<0.01;BaxmRNA的相对表达量明显升高,P>0.05;与对照组相比,Bax/Bcl-2mRNA的比值显著升高,P<0.05。肿瘤细胞增殖能力明显受到抑制,P<0.01;并出现G1期阻滞和凋亡细胞百分率增加。结论:反义HMGB1基因转染能抑制胰腺癌细胞增殖,促进细胞凋亡,是胰腺癌基因治疗的策略之一。OBJECTIVE： To study the apoptosis induction effect and Bcl-2/Bax mRNA expression changes of human antisense high mobility group box 1 gene on human pancreatic adenocarcinoma cell line （panc1）. METHODS： HMGB1 was cloned and reversely inserted into eukaryotic expression vector pcDNA3. 1 to get pcDNA3. 1/anti-HMGB1, which was then transfected into pancl cells by liposome method. The positive cell clones were abtained by G418 selection, and HMGB1, Bcl-2 and Bax mRNA expression were detected by RT-PCR. The cell proliferating activity in vitro was examined by MTT analysis, and the cell cycle and cell apoptosis were assessed by flow cytometry. RESULTS： The pcDNA3. 1/anti HMGB1 vector was obtained, pcDNA3. 1/anti- HMGB1 transfection could inhibit HMGB1 and Bcl-2 gene expressions in pancl cells, otherwise enhance the Bax mRNA expression. The proliferation of cultured pancreatic adenocarcinoma cells was suppressed significantly（P〈0.01）. The cell cycle G1 phase arrest appeared and apoptosis cell number obvious increased. CONCLUSIONS Antisense HMGB1 gene can inhibit the proliferation of human pancreatic adenocarcinoma cells and promote the cell apoptosis, and may be used as one of the target genes of pancreatic adenocarcinoma gene therapy.

关 键 词：高迁移率族蛋白质类 胰腺肿瘤 细胞凋亡 

分 类 号：R735.9[医药卫生—肿瘤]