尾型同源盒基因2基因表达对胃癌细胞生长影响的研究

目的 观察尾型同源盒基因2(Cdx2)基因过表达对胃癌细胞生物学性状的影响.方法 构建人Cdx2真核表达载体,并转染胃癌细胞MGC-803.实验分为四组:未转染组、转染pCMVHA组、转染pCMV-GAPDH-HA组、转染pCMV-Cdx2-HA组.应用荧光定量PCR检测各组Cdx2基因的表达情况,Western blot法检测各组Cdx2蛋白的表达情况,MTT法观测细胞增殖,透射电镜观察转染细胞形态学的改变,流式细胞仪分析细胞周期和凋亡.结果 成功构建了pCMV-Cdx2-HA真核表达载体,在瞬时转染pCMV-Cdx2-HA质粒48 h的MGC-803细胞有较高水平的Cdx2基因mRNA和蛋白的表达;与未转染组、转染pCMV-HA组、转染pCMV-GAPDH-HA组比较,转染pCMV-Cdx2-HA组的胃癌细胞在转染72 h时生长受到明显抑制,G0/G1期比例上升,S期比例下降,胃癌细胞的凋亡率也明显升高(P＜0.05);转染pCMV-Cdx2-HA的胃癌细胞胞核消失,核染色质固缩、解体,内质网、高尔基复合体膨大成泡状.结论 Cdx2基因可明显抑制胃癌细胞的生长,提示Cdx2参与了胃癌的生长调控.

Caudal-related homeobox 2 gene expression and its effects on human gastric carcinoma cell line

Objective To investigate the effect of human caudal-related homeobox 2(Cdx2)gene expression on human gastric carcinoma cell line MGC-803.Methods pCMV-Cdx2-HA eukaryotic expression plasmid was constructed by using DNA recombinant method.The MGC-803 cells were divided into 4 groups:nontranfected,transfected with pCMV-HA,transfected with pCMV-GAPDH-HA,transfected with pCMV-Cdx2-HA. Cdx2 gene and its protein expression was detected by quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot techniques respectively.The effects of Cdx2 overexpression on the growth of MGC-803 cells in vitro was assessed by measuring MTT,flow cytometry and transmission electron microscopy analysis.Results In MGC-803 cells transfected with pCMV-Cdx2-HA,cell proliferation was significantly suppressed,the cells was ultrastructurally destroyed,cell cycle progression was blocked in G0/G1 and apoptosis rate was higher(P＜0.05)in comparison with nontransfected cells or cells transfected with empty vector.Conclusion It indieated that Cdx2 gene can suppress the growth of gastric carcinoma and may save as a novel therapeutic target.