摘要
目的应用基因芯片技术筛选人子宫内膜癌细胞株中获得性紫杉醇(TAX)耐药相关基因。方法采用大剂量间歇诱导法,用TAX反复冲击诱导培养人子宫内膜癌Ishikawa细胞株,建成Ishikawa/TAX耐药株。应用Affymetrix Human Genome U133 Plus2.0表达谱芯片检测Ishikawa/TAX与Ishikawa细胞株基因。通过Gene Ontology(GO)聚类分析对差异表达的基因予以分类。采用qRT-PCR对部分差异表达的基因进行验证。结果与Ishikawa细胞相比,Ishikawa/TAX细胞中有110条基因表达有显著性差异(ratio比值>3),包括87个上调基因、23个下调基因,GO分类涉及细胞信号传导和调控、细胞黏附分子、细胞代谢及转录调控等;其中上调最明显的基因为S100A12,其次有CYP1B1、FUBP1、CEACAM6,下调基因有TNFAIP3、CD44、DKK1等。qRT-PCR检测S100A12、CYP1B1在耐药细胞中高表达,TNFAIP3、CD44在耐药细胞中低表达,与基因芯片结果一致。结论采用基因芯片技术筛选出的人子宫内膜癌获得性TAX耐药相关基因的上调基因有S100A12、CYP1B1、FUBP1、CEACAM6,下调基因有TNFAIP3、CD44、DKK1。
Objective To screen and identify the acquired taxol ( TAX)-resistant related genes in human endometrial carcinoma cell line by applying gene chip technology.Methods We established the Ishikawa/taxol ( TAX)-resistant cell line by intermittent and repeated exposure to TAX of a high concentration on the human endometrial carcinoma cell line Ish-ikawa.The genes of Ishikawa/TAX and Ishikawa cells were examined by using Affymetrix Human Genome U133 Plus2.0 expression profile chip.The differentially expressed genes were classified by Gene Ontology ( GO) cluster analysis.Real-time quantitative PCR ( qRT-PCR) was performed to confirm part of the differentially expressed genes.Results Compared with Ishikawa cells, there were 110 significantly differential expression genes in Ishikawa/TAX cells (ratio〉3), of which the up-regulated and down-regulated genes were 87 and 23, respectively.The classification of GO involved the cell signal transduction and regulation, cell adhesion molecule, cell metabolism and transcription regulation, etc.S100A12 was up-regulated significantly, followed by CYP1B1, FUBP1 and CEACAM6.TNFAIP3, CD44 and DKK1 were significantly down-regulated.The qRT-PCR showed that S100A12 and CYP1B1 was highly expressed in the drug-resistant cells, and TN-FAIP3 and CD44 was lowly expressed in the drug-resistant cells, which coincided well with the results of cDNA microarray. Conclusion Gene chip technology screens that the up-regulated genes of acquired TAX-resistant related genes in human endometrial carcinoma cell line are S100A12, CYP1B1, FUBP1 and CEACAM6, and the down-regulated genes are TN-FAIP3, CD44 and DKK1.
出处
《山东医药》
CAS
北大核心
2015年第19期21-24,共4页
Shandong Medical Journal
关键词
基因芯片技术
子宫肿瘤
子宫内膜癌
紫杉醇
耐药基因
gene chip technology
uterine neoplasms
endometrial carcinoma
paclitaxel
drug-resistant gene