E2F-1基因过表达对胃癌细胞动力学影响机制  

Effects of E2F-1 gene overexpression on gastric cancer cell dynamics

作　　者：王连振[1] 李雷[1] 王长青[1] 解乃昌[1] 肖强[1] 谢玉波[2] 

机构地区：[1]广西医科大学第一附属医院胃肠腺体外科,广西壮族自治区南宁市530021 [2]广西医科大学第一附属医院麻醉科,广西壮族自治区南宁市530021

出　　处：《世界华人消化杂志》2009年第11期1069-1073,共5页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30860273;广西医药卫生重点基金资助项目;No.桂卫重200844~~

摘　　要：目的:了解E2F-1过表达对胃癌细胞MGC-803增殖、生长和细胞周期的影响,并探讨E2F-1对胃癌细胞动力学影响的分子机制.方法:对稳定转染E2F-1的胃癌MGC-803/E2F-1细胞(实验组)、转染空载体的MGC-803/EV(阴性对照组)及未转染的MGC-803细胞进行MTT法检测,绘制生长曲线和计算细胞增殖率;流式细胞仪检测各组细胞周期分布.提取各组细胞的总RNA,逆转录成cDNA,用荧光探针与含有21522条人类基因表达谱芯片进行杂交.采用LuxScan10K/A双通道激光扫描仪扫描芯片上两种荧光信号,应用LuxScan3.0图像分析软件对芯片图像进行处理和分析.结果:与两个对照组相比,胃癌MGC-803/E2F-1细胞的生长受到明显抑制,增殖率下降(26.61%±5.19%vs93.4%±10.29%,100%±0.00%,均P<0.05);细胞周期阻滞在G0/G1期.在检测的21522条基因中,与细胞增殖、生长和细胞周期相关的差异性表达基因有20条,上调基因9条,下调基因11条.结论:E2F-1基因过表达抑制胃癌细胞的增殖和生长,细胞周期停滞在G0/G1期,其分子机制可能与筛选出来的20条差异表达基因有直接的关系.AIM： To understand the effect of E2F-1’s over-expression on gastric cancer cell proliferation, growth and cell cycle and to investigate the molecular mechanism of E2F-1’s influence on gastric cancer cell dynamics.METHODS： The MGC-803/ E2F-1 （experimental group） gastric cancer cells which were stably transfected with E2F-1, and containing unpossessed carriers （negative control group）, and the cells without transfection were performed MTT test to draw growing curve, and to compute cell proliferation rate. Cell cycles were examined by flow cytometry. The total RNA was extracted, and the cDNA was obtained using reverse transcription polymerase chain reaction （RT-PCR）, and then the fluorescent probes were hybridized with gene chip containing 21 522 human 22K gene expression profiles. Subsequently, the two signal images were scanned by Lux Scan 10K/A dual pathways laser scanner and analyzed by LuxScan 3.0 image analysis software.RESULTS： Compared with two control groups, the growth of the MGC-803 gastric cancer cells stably transfected with E2F-1 was obviously inhibited, and proliferation rate was decreased （26.61% ± 5.19% vs 93.4% ± 10.29%, 100% ± 0.00%, both P 〈 0.05）, cell cycle staged in periods of G0/G1. Of the 21 522 target genes, there were 20 cell proliferation, growth and cell cycle-related differential expression ones. Of the 20 genes, 9 were up-regulated and 11 were down-regulated. CONCLUSION： E2F-1 overexpression inhibits gastric cancer cell proliferation, growth and cell cycle, then cell cycle remains in periods of G0/G1, and perhaps the molecular mechanism has direct relationship with the 20 selected differential expression genes.

关 键 词：胃癌 E2F-1 基因表达谱芯片 差异表达基因 

分 类 号：R735.2[医药卫生—肿瘤] R737.9[医药卫生—临床医学]