摘要
目的:分析胃癌组织与癌旁正常胃黏膜组织中miRNA的差异表达情况。方法:收集胃癌组织和其相应癌旁正常胃黏膜组织共33对,经抽提、纯化RNA后,反转录合成荧光分子Hy3的cDNA探针,将其与miRCURYTMLNA Array(v.16.0)(Exiqon公司)芯片进行杂交,应用Axon GenePix 4000B芯片扫描仪来扫描miRNA芯片的荧光强度,GenePix Pro 6.0软件(Axon)把图像转化为数字信号,以差异大于2倍的为标准来确定胃癌黏膜组织中差异表达的miRNA。再用实时荧光定量PCR方法验证芯片结果中表达异常增高的miR-105在33例胃癌组织中的表达情况。结果:miRNA表达谱芯片结果显示:胃癌组织共中有51种miRNA表达异常,其中miR-105、miR-548n、miR-214*、miR-4309等31种miRNA表达上调,miR-31、miR-1275、miR-26b*、miR-744等20种miRNA表达下调;实时荧光定量PCR证实与癌旁正常胃黏膜组织相比,胃癌组织中miR-105表达显著上调(P<0.01)。结论:miR-105在胃癌组织的表达明显高于正常胃黏膜组织,可能与胃癌的发生、发展相关。我们的研究为胃癌的发病机制和诊断治疗提供了一个新的研究方向。
Objective： To investigate the miRNA expression difference between gastric cancer and nornmal gastric mucosa. Methods： Total miRNA extracted and purified from 33 pairs samples of gastric carcinoma＇s specimens and corresponding normal gastric tissue was tagged by fluorescent Hy3. The slides were scanned using the Axon GenePix 4000B microarray scanner.Scanned images were then imported into GenePix Pro 6.0 software （Axon） for grid alignment and data extraction, miRCURYTM LNA Array （v. 16.0 ） （Exiqon company）was used to detect differential expression of miRNA with the sdandard of 2-fold diffrential expression. Real-time fluorescence quantitative PCR was aapplied to verify miR-105 of miRNA array results. Results： The expression of 51 miRNA was diffrent between gastric cancer and nommal gastric mucosa.31 of them was up-regulated including miR-105, miR-548n, miR-214＊, miR-4309 and etc, 20 of them was down-regulated including miR-31, miR-1275, miR-26b＊, miR-744 and etc. The result of real-time fluorescence quantita- tive PCR shows that miR-105 highly expressed in gastric cancer, and the change has great statistical significance （P〈0.01）. Conclusions： Over expression of miRNA-105 in gastric carcinoma may be related with the pathogenesis and development of gastric cancer. Our study applied a new direction of the pathogenesis,diagnosis and treatment of gastric cancer.
出处
《现代生物医学进展》
CAS
2014年第16期3069-3072,共4页
Progress in Modern Biomedicine
基金
黑龙江省自然科学基金项目(D201120)
