乳腺癌MDA-MB-231细胞中Rab7a基因的网络分析  

Network analysis on Rab7a gene in breast cancer MDA-MB-231 cells

作　　者：徐凤英 刘儒 马丽 曲一帆 肖伟利 王玉珍[3] 谢基明 XU Fengying;LIU Ru;MA Li;QU Yifan;XIAO Weili;WANG Yuzhen;XIE Jiming(Graduate School, Inner Mongolia Medical University, Hohhot 010110,China;Department of Clinical Laboratory, People’ s Hospital, Inner Mongolia Autonomons Region, Hohhot 010010,China;College of Life Science, Inner Mongolia Agricultural University,Hohhot 010018,China)

机构地区：[1]内蒙古医科大学研究生学院,内蒙古呼和浩特010110 [2]内蒙古自治区人民医院检验科,内蒙古呼和浩特010010 [3]内蒙古农业大学生命科学学院,内蒙古呼和浩特010018

出　　处：《吉林大学学报（医学版）》2020年第3期464-469,共6页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81360394,31270922)。

摘　　要：目的:探讨三阴性乳腺癌(TNBC)MDA-MB-231细胞中Rab7a基因敲除后相关基因表达的变化,阐明Rab7a相关基因在TNBC中的作用。方法:选取对数生长期的乳腺癌ZR-75-30、MCF-7、T-47D、MDA-MB-231和HCC-1937细胞,采用Western blotting法检测乳腺癌ZR-75-30、MCF-7、T-47D、MDA-MB-231和HCC-1937细胞中Rab7a蛋白表达水平。慢病毒敲除TNBC MDA-MB-231细胞中Rab7a基因,分为阴性对照组和Rab7a基因敲除组,根据敲除的4种不同Rab7a序列分为KD1组、KD2组、KD3组和KD4组。采用qPCR法检测各组MDA-MB-231细胞中Rab7a基因敲除效率,全基因表达谱芯片检测Rab7a基因敲除效率最高组(KD2组)和阴性对照组差异基因,一体化通路分析(IPA)软件分析Rab7a基因与其他基因的相互作用。结果:在TNBC MDA-MB-231细胞中可见Rab7a蛋白表达。与阴性对照组比较,KD2组乳腺癌MDA-MB-231细胞中Rab7a基因敲除效率最高(P<0.01);与阴性对照组比较,KD2组乳腺癌MDA-MB-231细胞中有634个差异基因,其中上调基因和下调基因数均增多(P<0.01);差异基因分析,Rab7a基因敲除与癌症、细胞存活、细胞周期和细胞生长及转移等有密切联系;Rab7a的IPA网络图中eIF4F、IRS1和RPS6KB1表达下调,PRKAA1、IKBKE、VEGFA和转录因子ATF2表达上调。结论:Rab7a基因在TNBC MDA-MB-231细胞中以基因网络的形式发挥作用;Rab7a基因与多基因相互作用,参与癌细胞的病理过程。Objective:To investigate the changes of expressions of the related genes in the triple-negative breast cancer(TNBC)MDA-MB-231 cells after knockout of Rab7a gene,and to elucidate the roles of Rab7a-related genes in TNBC.Methods:The breast cancer ZR-75-30,MCF-7,T-47D,MDA-MB-231 and HCC-1937 cells in the logarithmic phase were selected.The expression levels of Rab7a protein in the breast cancer ZR-75-30,MCF-7,T-47D,MDA-MB-231,and HCC-1937 cells were detected by Western blotting method.The Rab7a gene in the MDA-MB-231 cells was knockout with lentivirus and the cells were divided into negative control group and Rab7a knockout group.According to the four different knockout Rab7a sequences,the Rab7a knockout group was divided into KD1 group,KD2 group,KD3 group and KD4 group.The knockout efficiencies of Rab7a gene in the MDA-MB-231 cells in various groups were detected by qPCR method,the differential genes in the highest knockout efficiency group(KD2 group)and negative control group were detected by full gene expression microarray,and the interaction between Rab7a gene and other genes was analyzed by integrated pathway analysis(IPA)software.Results:The Rab7a protein was expressed in the TNBC MDA-MB-231 cells.Compared with negative control group,the knockout efficiency of Rab7a gene in the MDA-MB-231 cells in KD2 group was the highest(P<0.01);compared with negative control group,there were 634 differential genes in the MDA-MB-231 cells in KD2 group,the number of up-regulated genes and down-regulated genes were increased(P<0.01);the differential gene analysis results showed that Rab7a knockout had the relationships with cancer,cell survival,cell cycle,cell growth,and migration.The expressions of eIF4F,IRS1 and RPS6KB1 in the Rab7a IPA network diagram were down-regulated,and the expressions of PRKAA1,IKBKE,VEGFA,and transcription factor ATF2 were up-regulated.Conclusion:Rab7a gene plays a role as a gene network in the TNBC MDA-MB-231 cells.The Rab7a gene interacts with the multiple genes and participates in the pathological

关 键 词：乳腺肿瘤 Rab7a 三阴性乳腺癌 MDA-MB-231细胞 一体化通路分析 

分 类 号：R737.9[医药卫生—肿瘤]