應(yīng)用cDNA芯片篩選膀胱移行細(xì)胞癌差異表達(dá)基因及驗(yàn)證研究
發(fā)布時(shí)間：2018-12-05 19:01
【摘要】：目的 應(yīng)用高通量cDNA芯片檢測(cè)膀胱移行細(xì)胞癌(BTCC)與正常膀胱黏膜組織基因表達(dá)譜,篩選共同差異表達(dá)基因,初步探討B(tài)TCC發(fā)生發(fā)展的分子機(jī)制,以期發(fā)掘潛在腫瘤特異基因。 方法 1.采用高通量cDNA芯片對(duì)6例的BTCC及3例癌旁正常膀胱黏膜組織基因表達(dá)譜進(jìn)行檢測(cè),篩選共同差異表達(dá)基因,進(jìn)而將其導(dǎo)入博奧生物分子功能注釋系統(tǒng)MAS3.0進(jìn)行GO功能分類及pathway生物信息學(xué)分析。 2.采用實(shí)時(shí)定量RT-PCR對(duì)4個(gè)顯著差異基因的表達(dá)進(jìn)行檢測(cè),對(duì)基因芯片結(jié)果的可靠性進(jìn)行驗(yàn)證。 結(jié)果 1.芯片結(jié)果共篩選出263個(gè)膀胱移行細(xì)胞癌共同差異表達(dá)基因,其中表達(dá)上調(diào)2倍以上的基因有92個(gè),表達(dá)下調(diào)2倍以上的基因有171個(gè)。差異基因GO功能分類涉及DNA轉(zhuǎn)錄與轉(zhuǎn)錄調(diào)節(jié)、細(xì)胞分裂、細(xì)胞周期、蛋白代謝分解、細(xì)胞凋亡和抗細(xì)胞凋亡、細(xì)胞增值、免疫應(yīng)答、DNA復(fù)制與修復(fù)等。差異基因通路涉及細(xì)胞周期通路、DNA聚合通路、泛素介導(dǎo)的蛋白通路、錯(cuò)配修復(fù)通路、P53信號(hào)通路、氧化磷酸化通路、細(xì)胞因子及其受體相互作用通路、PPAR信號(hào)通路、TGF-β信號(hào)通路等。 2.實(shí)時(shí)定量RT-PCR結(jié)果顯示,四個(gè)基因TOP2A、CDK1、BIRC5A、CAV1的相對(duì)表達(dá)量分別為10.45、5.35、28.46、0.14倍,與基因芯片結(jié)果的總體趨勢(shì)基本一致。 結(jié)論 1.基因芯片篩選出大量差異表達(dá)基因,通過(guò)GO功能富集和通路富集分析,充分證明BTCC的發(fā)生是一個(gè)涉及多基因、多步驟、多途徑調(diào)控的復(fù)雜過(guò)程。實(shí)驗(yàn)所篩選的差異基因表達(dá)譜及腫瘤相關(guān)基因?qū)Π螂装┑脑\斷具有重要的意義。 2.采用實(shí)時(shí)熒光定量PCR對(duì)芯片結(jié)果中TOP2A、CDK1、BIRC5A、CAV1基因的表達(dá)進(jìn)行檢測(cè),進(jìn)一步驗(yàn)證芯片結(jié)果的可靠性,并發(fā)現(xiàn)這些基因有可能成為診斷BTCC的潛在的標(biāo)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.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.14
本文編號(hào)：2365312
[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.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.14
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