结直肠癌相关基因tgfbr13’utr micrornas位点但分析及其功能研究.docxVIP

西南医科大学硕士学位论文的影响。(4)采用SPSS 西南医科大学硕士学位论文 的影响。(4)采用SPSS 1 7．0对所获得数据进行统计学分析。实验所得肿瘤 细胞Real-time qPCR结果比较采用独立样本t检验(Independent Samples T test)，方差不齐时采用近似t检验，P0．05为差异有统计学意义。 结果：(1)本论文筛选出与结直肠癌相关基因TGFBRl 3UTR单核苷 酸多态位点m334348，rs334349特异性结合且可信程度较高的miRNA为 miR-133b和miR-204．5p。(2)从5种结直肠癌细胞株中检测到具有目标SNP 的结直肠癌细胞株的是HT29。(3)将候选miRNA转入HT29结直肠癌细胞 株后，Real．time qPCR结果表明：与miR mimics／inhibitors control组相比， hsa-miR-133b mimics与hsa．miR-204．5p mimics都能显著降低HT29细胞中 TGFBRl mRNA的表达水平；而hsa．miR-1 33b inhibitors与hsa．miR．204．5p inhibitors转染后的HT29细胞中mRNA的表达水平都有显著升高。(4) Western．blot结果表明：与miR mimics／inhibitors control组相比，hsa-miR ．1 33b mimics和hsa．miR-204．5p mimics转染后的HT29细胞中TGFBRl蛋 白的表达水平降低，而hsa．miR-1 3 3 b inhibitors和hsa．miR-204．5p inhibitors 转染后的HT29细胞中TGFBRl蛋白的表达水平升高，说明干扰效果显著。 结论：利用生物信息学分析成功分析得到能与结直肠癌相关TGFBRl 3’UTR单核苷酸多态位点rs334348，rs334349特异性结合的miRNA为 miR一133b和miR-204．5p，实验结果证实miR-133b，miR-204．5p能特异性降 低TGFBRl mRNA和TGFBRl蛋白的表达。 关键词：结直肠癌；TGFBRl；3UTR；miRNA；单核苷酸多态性 万方数据 西南医科大学硕士学位论文The 西南医科大学硕士学位论文 The analysis and functional studies of Colorectal cancer related gene TGFBRl 3 UTR microRNAs’binding site Abstract：Objective：Colorectal cancer(CRC)is one of gastrointestinal cancer seriously harm to human health．Nationwide，there were nearly one million new cases each year，the mortality rate ranked third in developed countries．In China，the morbidity and mortality of CRC increased significantly with younger tendencyin recent years．The incidence of CRC was a complicate process in which many genes and factors take part in and go through a lot of stage developments．In order to improve the sensitivity and specificity of clinical treatment of CRC，clarify the mechanisms of colorectal cancer,looking for CRC markers，optimize the treatment programs of CRC has always been focused．Currently,the reports of single nucleotide polymorphisms of tumor-related genes(Single Nucleotide Polymorph Sims，SNPs)and a epigenetic markers：tiny RNA(microRNAs，miRNAs)，which provide new perspectives for the molecular mechanisms of CRC development