应用基因芯片技术筛选曲古菌素A诱导肺腺癌耐药细胞株A549/CDDP差异表达的凋亡基因  

Microarray-based apoptosis gene screening technique in trichostatin A-induced drug-resisted lung cancer A549/CDDP cells

作　　者：汪亚君 张海涛[2] 熊禹真 刘彬[2] 伍俊 

机构地区：[1]广东医科大学附属医院呼吸疾病研究所,广东湛江524023 [2]广东医科大学生物化学与分子生物学教研室,广东湛江524023

出　　处：《解放军医学杂志》2016年第8期645-652,共8页Medical Journal of Chinese People's Liberation Army

基　　金：广东省社会发展领域科技计划(2014A020212292);广东医科大学博士启动基金(ZY1105;XB1108);国家自然科学基金青年基金(81400023)~~

摘　　要：目的应用基因芯片技术筛选曲古菌素A(TSA)处理顺铂耐药肺癌细胞株A549/CDDP后的凋亡相关基因的表达变化,筛选TSA治疗顺铂耐药肺癌的靶分子。方法 TSA处理A549/CDDP细胞24h,以未经TSA处理的A549/CDDP细胞作为对照组,提取两组细胞总RNA并反转录为c DNA,采用Nimble Gen全基因组表达芯片检测基因表达情况,以Nimble Scan 2.5软件结合GO分析比较TSA处理组与对照组基因表达谱的变化,从差异表达基因中筛选出与凋亡和增殖相关的基因。结果与对照组相比,共筛选到TSA上调的凋亡相关基因85个以及下调的细胞增殖和生长相关基因43个。GO分析发现,这些基因的分子功能主要是调节细胞凋亡和细胞对化学刺激的抵抗作用,以及参与细胞生长、增殖、细胞生物质量维持等生物过程。TSA不仅激活了线粒体凋亡通路,而且激活了死亡受体相关凋亡通路,下调了与细胞耐药相关的基因BAG3和ABCC2。结论 TSA改变了A549/CDDP细胞中凋亡和增殖基因的表达,这些基因可能在TSA治疗顺铂耐药肺癌中发挥了重要作用。Objective To detect the expression profile changes of apoptosis-related genes in trichostatin A（TSA）-induced drug-resisted lung cancer cells A549/CDDP by microarray, in order to screen the target genes in TSA treating cisplatin-resisted lung cancer. Methods A549/CDDP cells were treated by TSA for 24 hours. Total RNA was extracted and reversely transcribed into c DNA. Gene expression levels were detected by the Nimble Gen whole genome microarray. Differences of expression profiles between TSA-treated and control group were measured by Nimble Scan 2.5 software and GO analysis. Apoptosis and proliferation related genes were screened from the expression changed genes. Results Compared with the control group, 85 apoptosis-related genes were up-regulated and 43 growth or proliferation related genes were down-regulated in the TSA-treated group. GO analysis showed that the functions of these genes are mainly regulating apoptosis, cell resistance to chemical stimuli protein, as well as regulating cell growth, proliferation and the biological process of maintaining the cell biological quality. TSA-activated not only the mitochondrial apoptotic pathways, but also the death receptor related apoptosis pathway, and down-regulated the drug resistance related genes BAG3 and ABCC2. Conclusion TSA may cause the expression changes of apoptotic and proliferation genes in A549/CDDP cells, these genes may play a role in TSA treating cisplatin-resisted lung cancer.

关 键 词：曲古霉素A 基因芯片 肺肿瘤 抗药性 肿瘤 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤]