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  

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

作　　者：刘揆亮[1] 范慧娟[2] 吴静[1] 封国生[3] 林香春[1] 刘红[1] 

机构地区：[1]首都医科大学附属北京世纪坛医院消化内科,北京100038 [2]第四军医大学唐都医院消化内科,陕西西安710038 [3]首都医科大学附属北京朝阳医院肿瘤科,北京100020

出　　处：《基础医学与临床》2016年第6期794-798,共5页Basic and Clinical Medicine

基　　金：中国铁路总公司科技研究开发计划(J2014C011-1);北京市卫生系统高层次卫生技术人才培养专项(2011-RC1);北京市教育委员会科技发展计划(KM201210025015)

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。Objective To screen and verify the 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 were 15 altered miRNAsin group of early cancer; compared with group of normal control,there were 13 altered miRNAs in group of early cancer. Level ofmiR-204,miR-193 b 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-193 b was significantly lower in patients with adenomas,early cancer or advanced carcinoma（ P〈0. 05,P〈0. 01 and P〈0. 05 respectively）.miR-193 b 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-193 b are the potential novel biomarkers for early diagnosis of colorectal carcinoma. miR-193 b may have tumor suppressing effect to adenoma-adenocarcinoma sequence.

关 键 词：微小RNA miR-193b miR-204 结直肠癌 结直肠腺瘤 

分 类 号：R735.34[医药卫生—肿瘤]