基础医学与临床 ›› 2016, Vol. 36 ›› Issue (6): 794-798.

• 研究论文 • 上一篇    下一篇

结直肠腺瘤-腺癌发展序列中差异表达miRNAs的筛选及初步验证

刘揆亮1,范慧娟2,吴静1,封国生3,林香春1,刘红1   

  1. 1. 首都医科大学附属北京世纪坛医院
    2. 第四军医大学唐都医院
    3. 北京朝阳医院
  • 收稿日期:2015-12-03 修回日期:2016-03-29 出版日期:2016-06-05 发布日期:2016-05-27
  • 通讯作者: 吴静 E-mail:wujing36@126.com
  • 基金资助:
    中国铁路总公司科技研究开发计划专项课题;北京市卫生系统高层次卫生技术人才培养资助专项;北京市教育委员会科技发展计划面上项目

Screening and verification of altered miRNAs in colorectal adenoma-adenocarcinoma sequence

  • Received:2015-12-03 Revised:2016-03-29 Online:2016-06-05 Published:2016-05-27

摘要: 目的 筛选及初步验证结直肠腺瘤-腺癌发展序列中差异表达的miRNA,初步探索其可能机制。方法 MicroRNA芯片检测15例结直肠腺瘤患者、3例结直肠早癌患者和3例健康对照者血清中差异表达的miRNA。在18例结直肠腺瘤,9例结直肠早癌,5例进展期癌及5例正常对照者中用实时荧光定量PCR对进行初步验证。结果 早癌组与腺瘤组比较,差异表达的miRNA有15个;早癌组与正常组比较,差异表达的miRNA有13个。选择miR-204、miR-193b与miR-150进行初步验证。进展期癌组血清miR-150表达显著低于正常对照组(P<0.05);早癌组、进展期癌组血清miR-204表达均显著低于正常对照组(分别为P<0.05,P<0.01)。腺瘤组、早癌组和进展期癌组血清miR-193b表达均显著低于正常对照组(分别为P<0.05、P<0.01、P<0.05)。miR-193b mimic转染HCT116细胞后显著抑制其增殖(P<0.05),促进其凋亡(P<0.001)。并可显著抑制K-ras与β-catenin蛋白的表达水平。结论 血清miR-204与miR-193b可能成为诊断结直肠癌的生物标志物,miR-193b可能在腺瘤-腺癌序列中发挥抑癌作用。

关键词: 微小RNA, miR-193b, miR-204, 结直肠癌, 结直肠腺瘤

Abstract: Objective To screen and verify differentially altered miRNAs in colorectal adenoma-adenocarcinoma sequence and investigate possible mechanism. Methods To screen altered miRNAs in 15 patients of colorectal adenomas, 3 patients of colorectal early cancer and 3 cases of normal control using Human MicroRNA Array. To verify the results in 18 patients of colorectal adenomas, 9 patients of colorectal early cancer and 5 cases of normal control using real-time PCR. Results Compared with group of adenomas, there are 15 altered miRNAsin group of early cancer;compared with group of normal control,there are 13 altered miRNAs in group of early cancer. Level of miR-204,miR-193b and miR-150 was verified further. Serum miR-150 was significantly lower in patients with adavanced carcinoma than normal control (P<0.05). Serum miR-204 was significantly lower in patients with early cancer or adavanced carcinoma (P<0.05 and P<0.01 respectively). Serum miR-193b was significantly lower in patients with adenomas, early cancer or advanced carcinoma ( P<0.05,P<0.01 and P<0.05 respectively). miR-193b mimic significantly inhibited proliferation of HCT116 cells (P<0.05)and promoted apoptosis (P<0.001). meanwhile inhibiting the expression of K-ras and β-catenin. Conclusions Serum miR-204 and miR-193b may serve as novel biomarkers in early diagnosis of colorectal carcinoma. miR-193b may exert tumor suppressing effect in adenoma-adenocarcinoma sequence.

Key words: micro RNA, miR-193b, miR-204, Colorectal cancer, Colorectal adenoma