微RNA-9对胃癌细胞内CDX2基因表达的影响  

Effect of miRNA-9 on CDX2 gene expression in gastric cancer cells

作　　者：周建萍 毛振彪[2] 吴信华[3] ZHOU Jianping;MAO Zhenbiao;WU Xinhua(Department of Emergency Medicine,Changhai Hospital of Shanghai,Shanghai（200433）,Chin;Department of Gastroenterology,Affiliated Hospital of Nantong University,Nantong（226001）,China;Department of Surgical Laboratory,Affiliated Hospital of Nantong University,Nantong（226001）,China)

机构地区：[1]海军军医大学附属长海医院急诊科,上海200433 [2]南通大学附属医院消化科,226001 [3]南通大学附属医院外科综合实验室,226001

出　　处：《国际消化病杂志》2018年第5期314-319,共6页International Journal of Digestive Diseases

摘　　要：目的探讨改变胃癌细胞中微RNA-9(miRNA-9)的表达水平对尾型同源核转录因子2(CDX2)表达的影响。方法采用生物信息学预测调控CDX2的目的 miRNA。采用脂质体法将miRNA-9类似物、miRNA-9抑制物和无关序列质粒瞬时转染胃癌细胞株AGS、MKN45,以无关序列作为阴性对照。将细胞分为miRNA-9类似物组、miRNA-9抑制物组、阴性对照组、空白对照组。采用实时荧光定量PCR(qRT-PCR)检测miRNA-9和CDX2mRNA的表达水平,采用免疫印迹实验(Western blot)检测CDX2蛋白表达。结果 CDX2mRNA 3′UTR和miRNA-9有高度保守的结合靶点。与空白对照组、阴性对照组相比,miRNA-9类似物组细胞中的CDX2蛋白表达显著降低,miRNA-9抑制物组的CDX2蛋白表达显著上调,而CDX2mRNA表达未发生显著变化。结论在胃癌细胞株AGS、MKN45中,miRNA-9具有抑制CDX2蛋白表达的作用,而对CDX2mRNA表达无影响。抑制miRNA-9表达后,CDX2蛋白表达明显上调。Objective This paper aims to investigate the effect of microRNA-9 （miRNA-9） expression on the expression of tail-type homologous nuclear factor 2 （CDX2） in gastric cancer cells.Methods The bioinformatics method was used to predict the target miRNA for CDX2 regulation. Using the liposome method, the miRNA-9 promoters, miRNA-9 inhibitors, and unrelated sequence plasmids were transiently transfected into gastric cancer cell lines AGS and MKN45. All cells were divided into four groups and designated as the miRNA-9 promoter transfection group, the miRNA-9 inhibitor transfection group, the negative control transfection group, and the blank control group. The qRT-PCR was used to detect the change expression of miRNA-9 and CDX2 mRNA. The western blot was used to detect the change expression levels of CDX2 protein.Results The bioinformatics analysis revealed that the human CDX2 mRNA 3′UTR contains perfect complementarity region with a seed sequence of miRNA-9. Compared with the blank control group and the negative control group, the expression of CDX2 protein in the miRNA-9 promoter group was significantly decreased. While the expression of CDX2 protein was significantly up-regulated in the miRNA-9 inhibitor group, the expression of CDX2 mRNA had no significant change.Conclusions In gastric cancer cell lines AGS and MKN45, miRNA-9 suppresses the expression of CDX2 protein, but it has no effect on CDX2 mRNA expression. After inhibiting miRNA-9, the expression level of CDX2 protein is up-regulated.

关 键 词：胃癌 miRNA-9 CDX2 转染 

分 类 号：R735.2[医药卫生—肿瘤]