IL27RA对EBV感染的鼻咽癌细胞TRAF1/2表达的调控及其生物学效应

Regulation on TRAF1/2 Expression and Biological Effect of IL27RA in NPC Cells with EBV

EB病毒(Epstein-Barr virus,EBV)感染是鼻咽癌(Nasopharyngeal carcinoma,NPC)最主要的危险因素,主要通过肿瘤坏死因子受体相关分子-1/2(Tumor necrosis factor receptor-associated factor-1/2,TRAF1/2)发挥促癌作用,为探究白介素27受体亚基α(Interleukin 27 receptorα,IL27RA)表达与NPC细胞EBV感染的关系及其生物学效应,本研究采用肿瘤基因组计划(The Cancer Genome Atlas,TCGA)数据库在NPC组织中筛选EBV感染相关信号分子TRAF1/2的潜在相互作用分子,培养携带EBV的NPC细胞株HK-1和C666-1及未携带EBV的NPC细胞株CNE-1及CNE-2,实时荧光定量PCR技术(RT-qPCR)及蛋白质印迹技术(Western Blot)检测IL27RA mRNA及蛋白表达,慢病毒介导细胞转染技术外源性干扰HK-1和C666-1细胞IL27RA的表达,CCK-8检测细胞增殖能力的变化情况,Transwell实验检测细胞侵袭能力的变化情况。结果显示,NPC组织中存在与TRAF1/2表达均显著正相关的分子IL27RA(相关系数R分别为0.61及0.69,P均小于0.001),同时IL27RA在NPC组织中高表达(P<0.001),HK-1和C666-1中IL27RA mRNA及蛋白的表达水平显著高于CNE-1及CNE-2细胞(P<0.01),干扰HK-1和C666-1细胞IL27RA表达后,TRAF1及TRAF2 mRNA及蛋白表达显著下调(P<0.01),且细胞增殖及转移能力均显著减弱(P<0.01)。本研究提示,干扰IL27RA表达可显著抑制携带EBV的NPC细胞TRAF1/2的表达及细胞增殖转移能力,IL27RA有望成为NPC新的分子靶标或标志物。

Epstein-Barr virus(EBV) infection is the most important risk factor of nasopharyngeal cancer(NPC),and it plays a role in promoting NPC through TNF receptor associated factor-1/2(TRAF1/2). To investigate the relationship between the expression of interleukin 27 receptor α(IL27RA)and EBV infection in NPC cells and biological effect of IL27RA,the Cancer Genome Atlas(TCGA)database was used to screen the potential interaction molecule of TRAF1/2,a signal molecule related to EBV infection. Culture HK-1 and C666-1(NPC cell lines carrying EBV)and CNE-1 and CNE-2(NPC cell lines without EBV). Real-time fluorescent quantitative PCR(RT-qPCR)and Western Blot were used to detect IL27RA mRNA and protein expression. Lenti-transfection technology was used to interfere with the expression of IL27RA in HK-1 and C666-1 cells exogenously. CCK-8 assay was used to detect the change of cell proliferation ability,and Transwell assay was used to detect the change of cell invasion ability. Results showed that IL27RA was positively correlated with the expression of TRAF1 and TRAF2 in NPC(the correlation coefficients R were0.61 and 0.69 respectively,P<0.001),and the expressions of IL27RA in NPC tissues were significantly higher than normal tissues(P<0.001). The mRNA and protein expression of IL27RA in HK-1 and C666-1 was significantly higher than that in CNE-1 and CNE-2(P<0.01). After interfering with the expression of IL27RA in HK-1 and C666-1 cells,the mRNA and protein expression of TRAF1 and TRAF2 were significantly upregulated(P<0.01). The ability of cell proliferation and metastasis were significantly weakened(P<0.01).The present study suggested that interference with IL27RA expression can significantly inhibit the expression of TRAF1/2 and cell proliferation and metastasis ability of NPC cells with EBV. IL27RA is expected to become a new molecular target or marker of NPC.