结肠癌微小RNA-126的表达情况及其与高尔基体磷酸化蛋白3的关系

Expression of microRNA 126 and its relation with Golgi phosphoprotein 3 in colon cancer

目的探讨结肠癌微小RNA-126(miR-126)的表达情况及其与高尔基体磷酸化蛋白3(GOLPH3)的关系。方法选取10例结肠癌及相应正常结肠上皮组织标本,检测miR-126的表达水平。将miR-126的模拟物和阴性对照(N.C.)分别转染至HCT116细胞,48 h后检测细胞中GOLPH3 mRNA和蛋白的表达情况。采用生物学信息方法预测miR-126和GOLPH3是否在存在靶向作用关系。构建p-GOLPH3 3'-UTR和p-GOLPH3 3'-UTR mut两种报告基因质粒,与miR-126模拟物进行共转染结肠癌细胞HCT116,同时以未转染的HCT116细胞为空白对照,仅转染N.C.的HCT116细胞为阴性对照组,检测各组细胞的荧光强度。结果结肠癌组织中miR-126的相对表达水平低于相应正常结肠上皮组织(P<0.05),转染miR-126模拟物的结肠癌细胞HCT116的GOLPH3 mRNA及蛋白表达水平均低于转染N.C.的结肠癌细胞HCT116。生物信息学分析结果提示miR-126与GOLPH3存在靶向作用关系。与空白对照组及阴性对照组比较,p-GOLPH3-3'UTR组的荧光值降低(P<0.05),但p-GOLPH3-3'UTR mut组的荧光值变化差异无统计学意义(P>0.05)。结论 miR-126在结肠癌组织中表达下调,其通过调控GOLPH3在结肠癌细胞中的表达可能是结肠癌的发病机制之一。

Objective To explore the expression of micro RNA126（miR-126） and its relation with Golgi phosphoprotein 3（GOLPH3） in colon cancer. Methods Ten specimens of colon cancer tissues and their corresponding normal colonic epithelial tissues were collected,then the miR-126 expression level was detected.miR-126 mimics and negative control（N.C.） were transfected into HCT116 cells separately,then the expressions of GOLPH3 mRNA and protein were detected in the cells after 48 hours.The target relationship between miR-126 and GOLPH3 was predicted by bioinformatics.Two reporter gene plasmids including p-GOLPH3 3′-UTR and p-GOLPH3 3′-UTR mut were constructed,then were co-transfected into HCT116 cells with miR-126 mimics.And HCT116 cells without transfection and transfetced by N.C. were enrolled as blank control group and negative control group respectively.The fluorescence intensity was detected in the cells of each group. Results The relative expression level of miR-126 in the colon cancer tissues was lower than that in the corresponding normal colonic epithelial tissues（ P 〈0.05）.The expression levels of GOLPH3 mRNA and protein were lower in the HCT116 cells transfected by miR-126 mimics compared to the cells transfected by N.C.The target relationship between miR-126 and GOLPH3 was found by bioinformatics.Compared to the blank control group and negative control group,the fluorescence intensity decreased in the p-GOLPH3- 3′UTR group（ P 〈0.05）,but no significant difference was observed in the fluorescence intensity changes of the p-GOLPH3-3′UTR mut group（ P 〉0.05）. Conclusion The expression of miR-126 is down-regulated in the colon cancer tissues.Regulation of GOLPH3 in the colon cancer cells is probably the mechanism of colorectal cancer.