吉林大学学报(医学版)

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上调 miR-21表达水平对胃癌 Kato Ⅲ 细胞中 CD44 mRNA 及蛋白表达的影响

蔡建英,杨效东,周明川   

  1. 重庆市第三人民医院普外科,重庆 400014
  • 收稿日期:2012-12-06 出版日期:2013-07-28 发布日期:2013-08-17
  • 通讯作者: 周明川(Tel:023-63511031,E-mail:mingchuanzhou120@126.com) E-mail:mingchuanzhou120@126.com
  • 作者简介:蔡建英 (1972-),女,四川省内江市人,主治医师,医学硕士,主要从事 胃癌干细胞相关研究。
  • 基金资助:

    重庆市卫生局医学科研计划项目资助课题 (06-2-028)

Influence of restoration of  miR-21 expression level in CD44 mRNAand  protein
 expressions in human gastric cancer Kato Ⅲ cells

CAI Jian-ying,YANG Xiao-dong,ZHOU Ming-chuan   

  1. Department of General Surgery,Third People’s Hospital of Chongqing City,Chon
    gqing 400014,China
  • Received:2012-12-06 Online:2013-07-28 Published:2013-08-17

摘要:

目的:探讨上调微小RNA-21( microRNA-21,miR-21) 表达对胃癌 Kato Ⅲ 细胞中肿瘤干细胞 (CSCs) 标志物 CD44 mRNA 及蛋白表达的影响,为胃癌细胞的恶性行为相关研究提供依据。方法:常规培养胃癌 Kato Ⅲ 细胞株,分为空白对照组和实验组,实验组又分为 Kato Ⅲ/miR-21 组和 Kato Ⅲ/control 组,分别予以 miR-21 模拟物 (hsa-miR-21 mimics) 或其阴性对照 (hsa-miR-21 control) 转染 Kato Ⅲ 细胞;通过荧光定量 RT-PCR 和 Western blotting法分别检测 CD44 mRNA 和蛋白在 Kato Ⅲ/control组 细胞和 Kato Ⅲ/miR-21组细胞中的表达水平。结果:miR-21 在 Kato Ⅲ/control 组细胞和 Kato Ⅲ/miR-21组细胞中的表达水平分别为 0.109 91±0.013 99 和 1.000 00±0.107 41,两者比较差异有统计学意义 (P<0.01);CD44 mRNA 在 Kato Ⅲ/control 组细胞和 Kato Ⅲ/miR-21 组细胞中的相对表达水平分别为 0.674 58±0.064 53 和 2.342 23±0.223 47,两者比较差异有统计学意义(P<0.01);CD44 蛋白在 Kato Ⅲ/control 组细胞和 Kato Ⅲ/miR-21组细胞中的相对水平分别为 0.045 929±0.007 396 和 0.176 048±0.009 900,两者比较差异有统计学意义(P<0.01)。结论:外源性上调 miR-21 表达水平能够诱导 Kato Ⅲ 细胞中 CD44 mRNA 及蛋白的过表达,可能诱导 CD44+胃癌 CSCs 的转化。

关键词:  , 微小RNA-21, CD44, 胃肿瘤, 肿瘤干细胞

Abstract:

Objective To investigate the influence of restoration of oncogenic microRNA-21 (miR-21) in CD44 mRNA and   protein expressions,one of the most common cancer stem cell (CSC) markers,in human gastric cancer Kato Ⅲ cells,and to  provide a new theoretical foundation for research on gastric cancer.Methods Human
gastric cancer Kato Ⅲ cells were cultured and divided into blank control group,Kato Ⅲ/miR-21 group and Kato Ⅲ/control group.Hsa-miR-21 mimics and  hsa-miR-21 control were transfected into Kato Ⅲ cells in Kato Ⅲ/miR-21 group and Kato Ⅲ/control group.Then the expressions of CD44 mRNA and protein in the abovecells were detected by  RT-PCR and Western blotting method.Results The relative expression levels of miR-21 in Kato Ⅲ/miR-21 cells was 1.000 00±0.107 41,which was significantly increased compared with Kato Ⅲ/control cells (0.109 91±0.013 99),there was significant difference( P<0.01). The mRNA level of CD44 in Kato /miR-21 cells was 2.342 23±0.223 47,which was significantly  increased compared with Kato Ⅲ/control cells (0.674 58±0.064 53),there was significant difference(P<0.01).The protein level of CD44 in Kato Ⅲ/miR-21 cells was 0.176 048±0.009 900,which was also significantly increased as compared with Kato Ⅲ/control cells (0.045 929±0.007 396),there was significant difference (P<0.01).Conclusion Restoration of miR-21 expression level can induce the over-xpressions of CD44 mRNA and protein in gastric cancer Kato Ⅲ cells,and may induce the transformation of gastric cancer CSCs.

Key words: microRNA-21, CD44, stomach neoplasms, cancer stem cells

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