山东医药
山東醫藥
산동의약
SHANDONG MEDICAL JOURNAL
2015年
19期
21-24
,共4页
基因芯片技术%子宫肿瘤%子宫内膜癌%紫杉醇%耐药基因
基因芯片技術%子宮腫瘤%子宮內膜癌%紫杉醇%耐藥基因
기인심편기술%자궁종류%자궁내막암%자삼순%내약기인
gene chip technology%uterine neoplasms%endometrial carcinoma%paclitaxel%drug-resistant gene
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。
目的：應用基因芯片技術篩選人子宮內膜癌細胞株中穫得性紫杉醇（ TAX）耐藥相關基因。方法採用大劑量間歇誘導法，用TAX反複遲擊誘導培養人子宮內膜癌Ishikawa細胞株，建成Ishikawa／TAX耐藥株。應用Affymetrix Human Genome U133 Plus2．0錶達譜芯片檢測Ishikawa／TAX與Ishikawa細胞株基因。通過Gene Ontology （ GO）聚類分析對差異錶達的基因予以分類。採用qRT-PCR對部分差異錶達的基因進行驗證。結果與Ishikawa細胞相比，Ishikawa／TAX細胞中有110條基因錶達有顯著性差異（ratio比值＞3），包括87箇上調基因、23箇下調基因，GO分類涉及細胞信號傳導和調控、細胞黏附分子、細胞代謝及轉錄調控等；其中上調最明顯的基因為S100A12，其次有 CYP1B1、FUBP1、CEACAM6，下調基因有 TNFAIP3、CD44、DKK1等。 qRT-PCR 檢測 S100A12、CYP1B1在耐藥細胞中高錶達，TNFAIP3、CD44在耐藥細胞中低錶達，與基因芯片結果一緻。結論採用基因芯片技術篩選齣的人子宮內膜癌穫得性TAX耐藥相關基因的上調基因有S100A12、CYP1B1、FUBP1、CEACAM6，下調基因有TNFAIP3、CD44、DKK1。
목적：응용기인심편기술사선인자궁내막암세포주중획득성자삼순（ TAX）내약상관기인。방법채용대제량간헐유도법，용TAX반복충격유도배양인자궁내막암Ishikawa세포주，건성Ishikawa／TAX내약주。응용Affymetrix Human Genome U133 Plus2．0표체보심편검측Ishikawa／TAX여Ishikawa세포주기인。통과Gene Ontology （ GO）취류분석대차이표체적기인여이분류。채용qRT-PCR대부분차이표체적기인진행험증。결과여Ishikawa세포상비，Ishikawa／TAX세포중유110조기인표체유현저성차이（ratio비치＞3），포괄87개상조기인、23개하조기인，GO분류섭급세포신호전도화조공、세포점부분자、세포대사급전록조공등；기중상조최명현적기인위S100A12，기차유 CYP1B1、FUBP1、CEACAM6，하조기인유 TNFAIP3、CD44、DKK1등。 qRT-PCR 검측 S100A12、CYP1B1재내약세포중고표체，TNFAIP3、CD44재내약세포중저표체，여기인심편결과일치。결론채용기인심편기술사선출적인자궁내막암획득성TAX내약상관기인적상조기인유S100A12、CYP1B1、FUBP1、CEACAM6，하조기인유TNFAIP3、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.
