应用cDNA芯片筛选膀胱移行细胞癌差异表达基因及验证研究

发布时间：2018-12-05 19:01

【摘要】：目的应用高通量cDNA芯片检测膀胱移行细胞癌(BTCC)与正常膀胱黏膜组织基因表达谱,筛选共同差异表达基因,初步探讨BTCC发生发展的分子机制,以期发掘潜在肿瘤特异基因。方法 1.采用高通量cDNA芯片对6例的BTCC及3例癌旁正常膀胱黏膜组织基因表达谱进行检测,筛选共同差异表达基因,进而将其导入博奥生物分子功能注释系统MAS3.0进行GO功能分类及pathway生物信息学分析。 2.采用实时定量RT-PCR对4个显著差异基因的表达进行检测,对基因芯片结果的可靠性进行验证。结果 1.芯片结果共筛选出263个膀胱移行细胞癌共同差异表达基因,其中表达上调2倍以上的基因有92个,表达下调2倍以上的基因有171个。差异基因GO功能分类涉及DNA转录与转录调节、细胞分裂、细胞周期、蛋白代谢分解、细胞凋亡和抗细胞凋亡、细胞增值、免疫应答、DNA复制与修复等。差异基因通路涉及细胞周期通路、DNA聚合通路、泛素介导的蛋白通路、错配修复通路、P53信号通路、氧化磷酸化通路、细胞因子及其受体相互作用通路、PPAR信号通路、TGF-β信号通路等。 2.实时定量RT-PCR结果显示,四个基因TOP2A、CDK1、BIRC5A、CAV1的相对表达量分别为10.45、5.35、28.46、0.14倍,与基因芯片结果的总体趋势基本一致。结论 1.基因芯片筛选出大量差异表达基因,通过GO功能富集和通路富集分析,充分证明BTCC的发生是一个涉及多基因、多步骤、多途径调控的复杂过程。实验所筛选的差异基因表达谱及肿瘤相关基因对膀胱癌的诊断具有重要的意义。 2.采用实时荧光定量PCR对芯片结果中TOP2A、CDK1、BIRC5A、CAV1基因的表达进行检测,进一步验证芯片结果的可靠性,并发现这些基因有可能成为诊断BTCC的潜在的标志物。
[Abstract]:Objective to detect the gene expression profiles of (BTCC) and normal bladder mucosa in bladder transitional cell carcinoma (TCC) by using high throughput cDNA microarray, and to screen common differentially expressed genes, and to explore the molecular mechanism of BTCC development. In order to explore the potential tumor specific genes. Method 1. High throughput cDNA microarray was used to detect gene expression profiles in 6 cases of BTCC and 3 cases of adjacent normal bladder mucosa to screen common differentially expressed genes. Then it was introduced into Boao Biomolecular function annotation system (MAS3.0) for GO functional classification and pathway bioinformatics analysis. 2. The expression of four differentially expressed genes was detected by real-time quantitative RT-PCR, and the reliability of the gene chip results was verified. Result 1. A total of 263 differentially expressed genes were screened in bladder transitional cell carcinoma. Among them, 92 genes were up-regulated more than 2 times, and 171 genes were down-regulated. Differential gene GO functional classification involves DNA transcription and transcription regulation, cell division, cell cycle, protein metabolism, apoptosis and anti-apoptosis, cell proliferation, immune response, DNA replication and repair. The differential gene pathway involves cell cycle pathway, DNA polymerization pathway, ubiquitin mediated protein pathway, mismatch repair pathway, p53 signal pathway, oxidative phosphorylation pathway, cytokine and its receptor interaction pathway, PPAR signaling pathway. TGF- 尾 signaling pathway. 2. The results of real-time quantitative RT-PCR showed that the relative expression levels of the four genes TOP2A,CDK1,BIRC5A,CAV1 were 10.455.35,28.46N 0.14 times, respectively, which were basically consistent with the general trend of gene chip results. Conclusion 1. A large number of differentially expressed genes were screened by gene microarray. Through the analysis of GO function enrichment and pathway enrichment, it is fully proved that the occurrence of BTCC is a complex process involving multi-gene, multi-step and multi-pathway regulation. The differential gene expression profiles and tumor-related genes screened by the experiment are of great significance in the diagnosis of bladder cancer. 2. Real-time fluorescence quantitative PCR was used to detect the expression of TOP2A,CDK1,BIRC5A,CAV1 genes in the microarray results. The reliability of the microarray results was further verified and it was found that these genes might be a potential marker for the diagnosis of BTCC.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.14

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