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Proline rich 11(PRR11)是本课题组鉴定的一个新的肿瘤相关基因。为研究PRR11介导肺癌发生发展相关的分子机制,本研究分析了PRR11表达被抑制后人肺癌细胞系H1299的全基因组基因表达谱的变化。首先,采用siRNA抑制H1299细胞中PRR11的表达,提取总RNA,采用基因芯片分析全基因组基因表达谱的变化。然后,对呈现差异表达的基因进行GO和Pathway富集分析,并对部分重要的候选基因进行定量RT-PCR验证。基因芯片结果表明,采用siRNA有效抑制H1299细胞中PRR11表达后,共有550个基因的mRNA水平出现明显变化,其中139个基因表达上调,411个基因表达下调。生物信息学分析结果表明,上述差异表达的基因显著富集于细胞周期和MAPK通路。定量RT-PCR验证分析结果表明,PRR11表达抑制后确实可导致多个与细胞周期和肿瘤发生发展密切相关的基因(包括DHRS2、EPB41L3、CCNA1、MAP4K4、RRM1、NFIB)呈现显著的表达变化。这些结果提示,PRR11可能通过上述通路和/或基因的表达变化参与肺癌的发生发展过程。
Proline rich 11 (PRR11) is a new tumor-related gene identified by our group. In order to study the molecular mechanisms involved in PRR11-mediated lung carcinogenesis, this study analyzed the genome-wide gene expression profiling of human lung cancer cell line H1299 after inhibition of PRR11 expression. First, the siRNA was used to inhibit the expression of PRR11 in H1299 cells and the total RNA was extracted. Gene chip analysis was used to analyze the genome-wide gene expression profiles. Then, the differentially expressed genes were analyzed by GO and Pathway enrichment, and some important candidate genes were confirmed by quantitative RT-PCR. Gene chip results showed that mRNA level of 550 genes was significantly changed after siRNA was used to effectively inhibit PRR11 expression in H1299 cells, of which 139 genes were up-regulated and 411 genes were down-regulated. Bioinformatics analysis showed that these differentially expressed genes were significantly enriched in the cell cycle and MAPK pathway. The results of quantitative RT-PCR analysis showed that PRR11 expression could indeed lead to significant expression changes of many genes (including DHRS2, EPB41L3, CCNA1, MAP4K4, RRM1, NFIB) closely related to cell cycle and tumorigenesis. These results suggest that PRR11 may participate in the development and progression of lung cancer through the above-mentioned changes in the expression of these pathways and / or genes.