维吾尔族结直肠癌组织中差异性表达的miRNA及其靶基因的研究

Study of differently expressed miRNA and the target gene in Uyghur nationality patients colorectal carcinoma tissue

目的:检测维吾尔族结直肠癌组织中差异性表达的miRNA及其靶基因,为维吾尔族结直肠癌发生机制的研究提供依据。方法:收集手术切除的维吾尔族结直肠癌患者的癌组织和维吾尔族正常结直肠黏膜组织标本,应用AFFX miRNA表达谱芯片技术筛查癌及正常组织中差异表达的miRNA并在miRNA靶基因预测数据库Target Scan与mi RDB中检测差异性表达的miRNA的靶基因。结果:hsa-mi R-196b,hsa-mi R-31和hsa-mi R-106a3种miRNA在维吾尔族结直肠癌组织中的表达较正常组织中呈高表达(<0.01)。HOXB8,PIK3C2A和CDH1分别为mi R-196b,mi R-31和mi R-106a的靶基因;HOXB8m RNA的表达水平在结直肠癌组织高于正常组织;mi R-196b mimics转染结果与其他对照组相比,转染24h和48h后,mi R-196b mimics组细胞HOXB8的表达均明显下调,表达量仅分别为对照组的45.03%(<0.05)和33.11%(<0.05);Western Blotting结果显示HOXB8蛋白在结直肠癌组织中表达低于正常结直肠粘膜(<0.05);Spearman检验分析结果显示,mi R-196b和HOXB8 m RNA的表达存在负相关(=0.005,=-0.783),而与HOXB8蛋白的表达无明显相关性(=0.294,=-0.194)。结论:在维吾尔族结直肠癌组织中hsa-mi R-196b,hsa-mi R-31和hsa-mi R-106a为高表达miRNA,HOXB8,PIK3C2A和CDH1分别为mi R-196b,mi R-31和mi R-106a的靶基因;mi R-196b和HOXB8 m RNA的表达存在负相关,而与HOXB8蛋白的表达无明显相关性。

Objective： To investigate the miRNA differently expression in Uyghur nationality patients＇ colorectal carcinoma tissue and predication of target gene of differently expressed miRNA,Study on mechanism for CHC will help to explain its development mechanism and provide a tool for detecting.Method： Uyghur nationality patients＇ colorectal carcinoma tissue and healthy tissue were collected and AFFX miRNA expression chip was used to determine the differendy expressed miRNAs.TargetScan and miRDB were used to predict the target gene of the differently expressed miRNAs.Results： The highest up-regulated miRNAs was hsa-miR-196b, hsa-miR-31 and hsa-miR-106a（P〈0.01）, HOXBS, PIK3C2A and CDH1 were the target gene of miR-196b, miR-31and miR-lO6a. The level of HOXB8 mRNA in colorectal carcinoma tissue was higher than that in healthy tissue.The results of miR-196b mimics transfection indicated that the level of HOXB8 mRNA significantly decreased after transfection 24h and 48h （P〈0.05）.The expression level of HOXB8 protein in colorectal carcinoma tissue was found to be significantly lower than that in healthy tissue （P〈0.05） by Western Blotting.Spearman testing showed that miR-196b was negatively correlated with HOXB8 mRNA （P=-0.005,r=-0.783）, while not correlated with HOXB8 protein （P=-0. 294,r=0.194）.Conclusion：We successfully detected the hsa-miR-196b, hsa-miR-31 and hsa-miR-106a were differently expressed miRNAs in Uyghur nationality paients＇ colorectal carcinoma tissue.HOXBS, PIK3C2A and CDH1 were the target gene of miR-196b,miR-31and miR-106a.respectively; HOXB8 was verified to be the target gene of miR-lO6a; miR-196b was negatively correlated with HOXB8 mRNA, while not correlated with HOXB8 protein.