尾型同源盒基因-2过表达对结肠癌细胞生物学性状的影响  

CDX2 gene overexpression and its inhibition on malignant progress of human colorectal cancer cell line

作　　者：李守帅[1] 孙学军[1] 郑见宝[1] 任海亮[1] 

机构地区：[1]西安交通大学医学院第一附属医院普通外科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2011年第1期114-118,127,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30672070;No.30400430);陕西省科学技术研究发展计划资助项目(No.2009K12-01)~~

摘　　要：目的观察尾型同源盒基因-2(CDX2)对结肠癌细胞生物学性状的影响,探讨CDX2基因在结肠癌发生发展中的作用。方法克隆人结肠CDX2基因,构建含CDX2的pEGFP-C1-CDX2表达载体,同时设pEGFP-C1空载体组及空白对照组。利用脂质体法将pEGFP-C1-CDX2和pEGFP-C1质粒分别转染Lovo细胞,应用Western blotting技术检测Lovo细胞中CDX2基因的表达;应用MTT法及流式细胞仪检测CDX2对Lovo细胞增殖、凋亡及周期的影响;应用ELISA法检测CDX2过表达对Lovo细胞基质金属蛋白酶-2(MMP-2)表达的影响。结果成功构建含CDX2真核表达载体。瞬时转染pEGFP-C1-CDX2的Lovo细胞48 h有较高水平的CDX2蛋白的表达;与pEGFP-C1空载体组及空白对照组比较,转染pEGFP-C1-CDX2组的Lovo细胞在转染72 h时生长未见明显抑制,G1期及S期所占比例无明显改变,凋亡率亦无明显增加(P均>0.05),但MMP-2分泌明显减少(P<0.05)。结论 CDX2过表达对Lovo细胞增殖、凋亡及周期无明显影响,但能够明显减弱Lovo细胞的侵袭力,从而对结肠癌浸润转移起抑制作用。Objective To investigate the effect of human caudal-related homeobox 2（CDX2） gene expression on human colorectal cancer cell line Lovo and explore CDX2＇s role in the pathogenesis and development of colorectal cancer.Methods Human colon gene CDX2 was cloned and the pEGFP-C1-CDX2 eukayotic expression plasmid was constructed by DNA recombinant method.Lovo cells were divided into three groups： transfected with pEGFP-C1-CDX2,transfected with pEGFP-C1,and non-transfected.We transfected pEGFP-C1-CDX2 and pEGFP-C1 plasmid into Lovo cells separately with lipofectamine 2000.CDX2 gene expression in Lovo cells was detected by Western blotting.The effects of CDX2 overexpression on the proliferation,apoptosis and cycle of Lovo cells were assessed by MTT and flow cytometry.The effects on the matrix metalloproteinase-2（MMP-2） expression were detected by ELISA method.Results pEGFP-C1-CDX2 eukayotic expression plasmid was constructed successfully and there was a higher expression of CDX2 protein in Lovo cells which had transient transfection of pEGFP-C1-CDX2 plasmid for 48 hours.Compared with that of the cells transfected with empty vector or non-transfected cells,the growth of cells transfected with pEGFP-C1-CDX2 was not inhibited obviously after 72 hours（P0.05）.The proportion of G1 and S phases of cell cycle progression did not change obviously（P0.05）.Apoptosis rate was not increased obviously,either（P0.05）.However,the secretion of MMP-2 was reduced obviously（P0.05）.Conclusion The overexpression of CDX2 gene has no obvious effect on the proliferation,apoptosis or cycle of Lovo cells,but it can significantly weaken the invasiveness and migration of Lovo cells;therefore,it may play an important role in inhibiting the invasion and metastasis of colorectal cancer.

关 键 词：尾型同源盒基因-2 结直肠癌 增殖 凋亡 周期 基质金属蛋白酶-2 

分 类 号：R735.3[医药卫生—肿瘤]