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.

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）