miR711靶向下调CD44抑制胃癌干细胞干性特征的研究

doi:10.3969/j.issn.1005-6483.2021.06.016

摘要/Abstract

摘要： 目的：探讨miR711对胃癌干细胞(gastric cancer stem cells,GCSCs)干性特征的影响及对CD44的靶向调控作用。方法：使用TCGA数据库验证miR711在胃癌组织以及正常胃黏膜组织中的表达水平。选取人胃癌干细胞CSCG和人胃癌细胞株MKN45，对CSCG细胞进行转染，分为miRNC组(转染miRNC)，miR711组(转染miR711mimics)及CSCG组(未转染CSCG)。实时荧光定量PCR检测各组细胞miR711、Bmi1、Sox2、Oct4、CD44的mRNA表达；Western Blot检测Bmi1、Sox2、Oct4、CD44蛋白表达；细胞肿瘤球形成实验检测细胞干性；生物信息预测miR711与CD44的靶向作用关系，荧光素酶实验验证。结果：TCGA数据分析显示，与正常胃黏膜组织比较，miR711在胃癌组织中的表达降低(P＜0.01)。miR711在miR711组细胞的水平高于miRNC组、CSCG组及MKN45组(P＜0.01)；miR711组细胞Bmi1、Sox2、Oct4、CD44 基因mRNA水平低于miRNC组、CSCG组(P＜0.01)；miR711组细胞Bmi1、Sox2、Oct4、CD44 蛋白水平低于miRNC组、CSCG组(P＜0.01)；培养1周、2周时，miR711组悬浮细胞球百分比低于CSCG组、miRNC组(P＜0.01)；miR711+WTCD44组细胞荧光素酶活性低于WTCD44组(P＜0.01)。结论：过表达miR711可抑制GCSCs干性，可能是通过下调CD44基因表达实现的。

关键词: miR711, CD44, 胃癌干细胞, 干性

Abstract: Objective：To investigate the effect of miR711 on the characteristics of stemness in gastric cancer stem cells(GCSCs) and its targeted regulation on CD44.ethods：We examined the expression of miR711 in gastric cancer tissues and normal gastric tissues using TCGA database.Human gastric cancer stem cell CSCG and human gastric cancer cell line MKN45 were selected.CSCG were transfected and divided into miRNC group(transfected with miRNC),miR711 group(transfected with miR711mimics) and CSCG group(no transfected).qRTPCR was used to detect the expression of miR711,Bmi1,Sox2,Oct4 and CD44 mRNA.Western blot was used to detect the protein expression of Bmi1,Sox2,Oct4,and CD44.Tumor ball formation test was used to detect the cell stemness.The target effect of miR711 and CD44 was confirmed by bioinformatics methods and the luciferase reporter assay.Results：The expression of miR711 was significantly lower in the gastric cancer tissues than in the normal gastric tissues(P＜0.01).The expression of miR711 in miR711 group was higher than that in miRNC group,CSCG group and MKN45 group(P＜0.01).The mRNA level of Bmi1,Sox2,Oct4 and CD44 in miR711 group were lower than those in miRNC group and CSCG group(P＜0.01).The expression level of Bmi1,Sox2,Oct4,CD44 in miR711 group were lower than those in miRNC group and CSCG group(P＜0.01).At 7 and 14 days,the percentage of suspended cell ball in miR711 group was lower than that in miRNC group and CSCG group(P＜0.01).Compared with WTCD44 group,the luciferase activity of miR711+WTCD44 group significantly decreased(P＜0.01).Conclusion：The overexpression of miR711 can inhibit the stemness of the GCSCs,which may be achieved by downregulating the expression of CD44 gene.

Key words: miR711, CD44, gastric cancer stem cells, stem

李建刚, 李亮, 帕合热迪尼·玉素莆等. miR711靶向下调CD44抑制胃癌干细胞干性特征的研究[J]. 临床外科杂志, 2021, 29(6): 554-558.

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