siRNA介导的PRR11表达抑制导致肺癌细胞系基因表达谱变化的分析  

Analysis of Gene Expression Profile Changes Responding to siRNAmediated PRR11 Depletion in Lung Cancer H1299 Cells

作　　者：龙银江[1,2] 吉颖[1,2] 翁华莉[1,2] 张春冬[1,2] 谢濛宇[1,2] 蔡伟[1,2] 王义涛[1,2] 朱远远[1,2] 李轶[1,2] 张莹[1,2] 卜友泉[1,2] 

机构地区：[1]重庆医科大学生物化学与分子生物学教研室,重庆400016 [2]重庆医科大学分子医学与肿瘤研究中心,重庆400016

出　　处：《中国细胞生物学学报》2013年第2期196-202,共7页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:30801356;81171879)资助的课题~~

摘　　要：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 novel cancer-associated gene identified by our group. In the present study, in order to clarifying the molecular mechanisms of PRR11 in lung cancer development, the gene expression profile changes responding to the siRNA-mediated PRR11 depletion was analyzed, siRNAs were used to inhibit the PRR11 expression, total RNAs were then prepared and subjected to microarray analysis. The differentially expressed genes were enriched for GO and pathway analysis, qRT-PCR was used to verify several important differentially expressed genes. Microarray analysis revealed that siRNA-mediated PRR11 depletion resulted into the expression changes of 550 genes including 139 upregulated and 411 downregulated. Bioinformatic analysis indicated that the 550 differentially expressed genes were mainly enriched in cell cycle and MAPK pathways, qRT-PCR analysis verified that siRNA-mediated PRR11 depletion led to the expression changes of several cell cycle- and cancer-related genes including DHRS2, EPB41L3, CCNA1, MAP4K4, RRM1 and NFIB. Taken together, the present study suggested that PRR11 might be implicated in lung cancer development via regulating the aforementioned genes and/or pathways.

关 键 词：PRR11 基因芯片 肺癌 差异表达基因 

分 类 号：R734.2[医药卫生—肿瘤]