通过数字基因表达谱和RNA测序技术鉴定自然杀伤／T细胞淋巴瘤的耐药相关基因  

Identification of drug - resistance related genes in gene expression and RNA -sequencing

作　　者：李鑫[1] 程雅颂 张旭东[1] 曹玲[1] 周志远[1] 孙振昌[1] 张明智[1] 

机构地区：[1]郑州大学第一附属医院肿瘤科,450052

出　　处：《中华实验外科杂志》2017年第10期1754-1756,共3页Chinese Journal of Experimental Surgery

基　　金：2014年河南省教育厅科技攻关计划项目（14A320089）;2014年河南省科技厅科技攻火项目（142102310315）

摘　　要：目的筛选正常自然杀伤（NK）细胞及NK／T细胞淋巴瘤细胞株YTS、SNK-6差异表达基因，明确耐药相关基因表达。方法通过数字基因表达谱（DGE）测序技术检测正常NK细胞和NK／T细胞淋巴瘤细胞株YTS和SNK-6的DGE谱，分析3种细胞株中的差异表达基因。SNK-6细胞株与正常NK细胞比较，高表达的基因数为3184个，低表达的基因数为2764个；YTS细胞株与正常NK细胞比较，高表达的基因数为3978个，低表达的基因数为2172个；SNK-6细胞株与YTS细胞株比较，低表达的基因数为2425个，高表达的基因数为1068个。通过实时荧光定量聚合酶链反应（FQ—PCR）检测常见耐药相关基因表达，建立正常NK细胞和两个NK／T细胞淋巴瘤细胞株的数字基因表达谱。结果构建正常人NK细胞和两个NK／T细胞淋巴瘤细胞株的数字基因表达谱文库，筛选3个细胞株之间的明显差异表达基因；耐药相关基因多药耐药蛋白5（ABCC5）和红白血病病毒致癌基因同系物4（ERBB4）在NK／T细胞淋巴瘤细胞株中表达升高，其中，ABCC5基因在YTS及SNK-6细胞株中的mRNA水平（3．694-0．11、5．22±1．66）明显高于正常NK细胞（1．00±0．00），差异有统计学意义（P=0．001、0．048）；ERBB4基因在YTS及SNK-6细胞株中的mRNA水平（6．87±0．56、14．79±1．84）明显高于正常NK细胞（1．00±0．00），差异有统计学意义（P=0．003、0．006）。结论ABCC5和ERBB4的高表达与NK／T细胞淋巴瘤的耐药性明显相关。Objective To explore the differentially expressed genes among normal natural killer （NK） cell and NK/T cell lymphoma cell lines TTS and SNK - 6 and define the expression of common drug - resistam：e related genes in NK/T cell lymphoma cell lines. Methods The digital gene expression （DGE） profiles of normal NK cells and NK/T -cell lymphoma cells YTS and SNK -6 were detected by DGE sequencing technology. The differential expressed genes among the three cell lines were also ana- lyzed. 3 184 genes were upregulated and 2 764 genes were downregulated in SNK - 6 cells as compared with normal NK cells. 3 978 genes were upregulated and 2 172 genes were downregulated in YTS cells as compared with normal NK cells. However, as compared with YTS cells, 1 068 genes were upregulated and 2 425 genes were downregulated in SNK - 6 cells. The expression levels of common drug - resistance relat- ed genes were examined by real - time flum＇escent quantitative polymerase chain reaction （ FQ - PCR）. Consequently, the DGE profiles of normal NK ceils and two NK/T cell lymphoma cell lines were estab- lished. Results DGE libraries of one normal NK cell line and two NK/T cell lymphoma cell lines were es- tablished and significantly differentially expressed genes of three cell lines were screened. The level of mul- tidrug resistance protein 5 （ ABCC5 ） mRNA was significantly higher in both YTS cells （ 3.69±0. 11, P = 0. 001 ） and SNK -6 cells （5.22 ± 1.66, P =0. 048） than in normal NK cells （1. 00±0. 00）. The level of erythroblastic leukemia viral oncogene homolog 4 （ERBB4） mRNA was significantly higher in both YTS cells （6.87± 0. 56, P = 0. 003 ） and SNK - 6 cells （ 14.79 ±1.84, P =0. 006） than in normal NK cells（ 1.00±0. 00）. Conclusion The highly expressed ABCC5 and ERBB4 may be associated with drug resistance in NK/T cell lymphoma. We can foresee that drug - resistance related genes might be targets for detection and treatment of NK/T cell lymphoma patients.

关 键 词：耐药基因 数字基因表达 NK/T细胞淋巴瘤 

分 类 号：R733.1[医药卫生—肿瘤]