胃肠富集Krüppel样因子基因转染对人胃癌细胞株SGC-7901及裸鼠移植瘤生长的影响

Effects of transfected gut-enriched Kriippel-like factor on growth of human gastric carcinoma cell line SGC-7901 and its xenograft in nude mice

目的探讨外源性胃肠富集Krueppel样因子（GKLF）基因转染对人胃癌细胞株SGC-7901在体内外的抗肿瘤效应。方法荧光实时定量PCR和Western印迹法检测GKLF基因转染前后人胃癌细胞株SGC-7901中GKLFmRNA和蛋白的表达。应用四甲基偶氮唑盐（MTT）法、流式细胞技术、克隆形成实验和细胞侵袭实验分别检测GKLF基因转染后SGC7901细胞增殖和侵袭力的变化。观察裸鼠移植瘤生长情况和应用免疫组织化学法检测移植瘤组织中微血管密度（MVD）。结果GKLF基因转染后，SGC7901-pcDNA3．1-GKLF组中GKLFmRNA（0．1216±0．0061）和蛋白（1．0547±0．0172）的表达明显高于SGC7901组[分别为（0．0029±0．0012）和（O．6240±0．0177）]和SGC7901pcDNA3．1组[分别为（O．0037±0．0013）和（O．5627±0．0510）]，P〈0．05。与SGC7901组和SGC7901-pcDNA3．1组相比，从48h开始，SGC7901-pcDNA3．1-GKLF组细胞生长速度减慢、细胞出现G0／G1期部分阻滞、克隆形成率低、细胞侵袭力明显减弱（P值均〈0．05）。SGC7901-pcDNA3．1-GKLF组皮下移植瘤生长速度减慢、瘤重轻、MVD低（P值均〈0．05）。结论GKLF基因转染可导致人胃癌细胞株SGC-7901的增殖活性和侵袭力降低，抑制裸鼠移植瘤生长和血管生成。

Objective To investigate the antitumour effects of transfected gut enriched Krtippellike factor（GKLF） on human gastric carcinoma cell line SGC-7901 in vitro and in vivo. Methods The expression of GKLF mRNA and protein in human gastric carcinoma cell line SGC-7901 were detected before and after transfeetion by real-time fluorescence quantitative PCR and Western blot, respectively. Proliferation and invasion in SGC-7901 were measured respectively by MTT assay, flow cytometry, colony formation assay and cell invasion assay after transfected with GKLF. The growth of xenograft was observed, the microvessel density（MVD） of xenograft tissue was determined by immunohistochemistry. Results The GKLF mRNA and protein in SGC-7901 were overexpressed after transfected with GKLF（P^0.05）. The proliferative speed of SGC7901-pcDNA3.1-GKLF group was markedly lower than that of SGC 7901 and SGC7901-pcDNA3.1 groups （P〈i0.05）. Transfected with GKLF caused part of the G0/G1 arrest, decreased clone formation rate and the invasion ability （P%0.05）. The growth speed of xenograft in SGC7901-pcDNA3. 1 GKLF group was lower, the weight and MVD of xenograft tissue in SGC7901-pcDNA3. 1-GKLF group were less （P^0. 05）. Conclusion Transfected with GKLF maysuppress proliferation and invasion in human gastric Carcinoma cell line SGC-7901, inhibit the growth and the angiogenesis of xenograft in nude mice.