IL-2、IFN-α、IFN-γ对肾透明细胞癌786-0细胞中B7-H4表达的影响  

Effect of IL-2,IFN-α and IFN-γ on expression of B7-H4 in clear cell renal cell carcinoma 786-0 cells

作　　者：徐一鹏[1] 朱绍兴[1] 刘维辉[2] 李永生[3] 陈贵平[1] 王华[1] 赵阳[1] 李方印[1] 王宗平[1] 

机构地区：[1]浙江省肿瘤医院泌尿外科,浙江杭州310022 [2]福建医科大学附属第二医院泌尿外科,福建泉州362000 [3]福建医科大学附属协和医院泌尿外科,福建福州350001

出　　处：《中国肿瘤生物治疗杂志》2013年第5期547-551,共5页Chinese Journal of Cancer Biotherapy

基　　金：浙江省卫生适宜技术成果转化项目(No.2013ZHB001);浙江省肿瘤医院优秀科研人才培育基金项目(No.2012YC004)~~

摘　　要：目的:探究IL-2、IFN-α和IFN-γ对人肾透明细胞癌786-0细胞B7-H4表达的影响。方法:IL-2、IFN-α、IFN-γ处理786-0细胞24 h后,RT-PCR法检测B7-H4 mRNA的表达,ELISA法、免疫细胞化学法、流式细胞术检测B7-H4蛋白的表达。结果:RT-PCR结果显示,IL-2组(0.75±0.06)、IFN-α组(0.68±0.05)、IFN-γ组(0.95±0.08)786-0细胞中B7-H4 mRNA的表达均明显高于未处理组细胞(0.30±0.03)(P<0.05)。免疫细胞化学染色结果显示,于786-0细胞膜与细胞质均可检测到B7-H4蛋白表达,IL-2、IFN-α、IFN-γ处理均可增加786-0细胞B7-H4蛋白的表达。ELISA结果显示,IL-2组[(44.89±0.97)ng/ml]、IFN-α组[(46.74±2.25)ng/ml]、IFN-γ组[(47.31±1.12)ng/ml]786-0细胞上清液中分泌型B7-H4的表达明显高于未处理组[(34.42±1.69)ng/ml](P<0.05)。流式细胞术检测结果表明,IL-2组[(44.89±0.94)%]、IFN-α组[(46.41±0.55)%]、IFN-γ组[(54.18±1.42)%]786-0细胞表面B7-H4蛋白的阳性表达率明显高于未处理组[(30.45±0.96)%](P<0.05)。结论:IL-2、IFN-α、IFN-γ在转录与翻译两个环节均可上调786-0细胞B7-H4的表达水平,其中以IFN-γ上调能力最强。Objective ： To explore the influence of IL-2, IFN-α and IFN-γ on the expression of B7-H4 in clear cell renal cell carcinoma 786-0 cells． Methods： Clear cell renal cell carcinoma 786-0 cells were stimulated by IL-2, IFN-α and IFN-γ for 24 h. The expression of B7-H4 mRNA was detected by RT-PCR. The expression of B7-H4 protein was detected by ELISA assay, cytoimmunochemistry assay and flow cytometry. Results： RT-PCR result showed that the expression of B7-H4 mRNA in IL-2 （0.75±0.06）, IFN-α （0.68±0.05） and IFN-γ （0.95±0.08） group cells was significantly higher than that in the untreated group （0.30±0.03） （P〈0.05）. Immunocytochemistry showed that the expression of B7-H4 protein was detected both in the cell membrane and cytoplasm, and the expression of B7-H4 protein was up-regulated after stimulated by IL-2, IFN-α and IFN-γ. ELISA result showed that the expression of soluble B7-H4 protein in the supernatants of IL-2 （44.89±0.97） ng/ml, IFN-α （46.74±2.25） ng/ml and IFN-γ group cells （47.31±1.12） ng/ml was significantly higher than that in the untreated group （34.42±1.69） ng/ml （P〈0.05）. Flow cytometry assay result showed that the positive expression rate of B7-H4 in IL-2 （44.89±0.94）%, IFN-α （46.41±0.55）% and IFN-γ （54.18±1.42）% group cells were significantly higher than that in the untreated group （30.45±0.96）% （P〈005）. Conclusion： IL-2, IFN-α and IFN-γ can up-regulate the expression of B7-H4, in which IFN-γ has the highest capacity.

关 键 词：肾透明细胞癌 B7-H4 IL-2 IFN-Α IFN-Γ 肿瘤免疫 

分 类 号：R692[医药卫生—泌尿科学]