基因芯片筛选结肠癌伊立替康耐药相关基因

Analysis of Gene Expression Patterns in an Irinotecan-resistance Colon Cancer Cell by cDNA Microarray

目的应用基因芯片技术进行结肠癌伊立替康耐药相关基因表达谱差异分析,探讨伊立替康结肠癌耐药发生机制。方法分别抽提人结肠癌细胞系SW480及其伊立替康耐药细胞系SW480/CPT总RNA,逆转录合成相应以Cy3标记的cRNA做探针,在Agilent基因芯片上杂交,Axon4000B扫描仪扫描芯片荧光信号图像,Genepix3.0图像软件对扫描图像进行数字化处理和分析。利用RT-PCR对筛选出的部分差异表达基因进行验证。结果筛选出差异表达基因1598个,其中基因表达上调911个,基因表达下调687个。谷胱甘肽S转移酶同工酶(GSTA)家族GSTA1、GSTA2、GSTA3、GSTA5明显上调,锌指蛋白(ZNF)家族多个成员也有显著差异表达。结论基因芯片结果提示,GSTA与ZNF基因可能在介导伊立替康耐药过程中起着重要的调控作用。

Objective To study the mechanism of irinotecan-resistant colon cancer by analyzing the differential gene expression patterns with cDNA microarray.Methods Total RNA was purified from irinotecan-sensitive colonic cancer cell line,SW480 and its irinotecan-resistance cell line,SW480/CPT.The cRNA retro-transcribed from RNA were labeled with Cy3 fluorescence as probes.The probes were hybridized with Agilent gene chips and the fluorescence images of the chips were obtained with Axon 4000B scanner as well as analyzed with Genepix 3.0 software.The microarray results were confirmed by reverse transcription-polymerase chain reaction.Results Of the 1598 genes with altered expressions,there were 911 up-regulated genes and 687 down-regulated genes.Glutathione S-Transferase（GST） isoenzyme family GSTA such as GSTA1,GSTA2,GSTA3 and GSTA5 were significantly up-regulated.The expression levels of many Zinc finger protein family members（ZNF） were also differentially regulated.Conclusion GSTA and ZNF subunit genes might play an important regulation role in the irinotecan resistance of colon cancer.