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目的:研究IL-2、IFN-α和IFN-γ对人肾透明细胞癌786-0细胞上B7-H1表达的影响。方法:IL-2、IFN-α和IFN-γ刺激786-0细胞,RT-PCR检测刺激后786-0细胞B7-H1mRNA的表达,免疫细胞化学、免疫荧光染色和流式细胞术检测刺激后786-0细胞中B7-H1蛋白的表达。结果:RT-PCR检测结果显示,IL-2、IFN-α和IFN-γ组786-0细胞的B7-H1mRNA表达量明显高于未经刺激的786-0细胞(0.75±0.06、0.68±0.05、0.95±0.08vs0.30±0.03,P(0.05或P<0.01);免疫细胞化学和免疫荧光检测结果显示,B7-H1蛋白主要表达在786-0细胞的细胞膜上,IL-2、IFN-α和IFN-γ刺激后B7-H1表达明显上调;流式细胞术检测结果显示,IL-2、IFN-α和IFN-γ组786-0细胞的B7-H1分子表达阳性率明显高于未经刺激的786-0细胞[(65.70±3.26)%、(56.52±1.75)%、(84.05±3.52)%vs(20.49±1.03)%,P<0.01]。结论:IL-2、IFN-α和IFN-γ均上调786-0细胞B7-H1mRNA和蛋白的表达,其中IFN-γ上调程度最高,IFN-α上调程度最低。
Objective: To investigate the effect of IL-2, IFN-α and IFN-γ on the expression of B7-H1 on human clear cell carcinoma 786-0 cells. Methods: The 786-0 cells were stimulated with IL-2, IFN-α and IFN-γ. The expression of B7-H1 mRNA in 786-0 cells was detected by RT-PCR. Immunocytochemistry, immunofluorescence staining and flow cytometry B7-H1 protein expression in 786-0 cells. Results: RT-PCR results showed that the expression of B7-H1 mRNA in 786-0 cells of IL-2, IFN-α and IFN-γ groups was significantly higher than that of 786-0 cells without stimulation (0.75 ± 0.06,0.68 ± 0.05 , 0.95 ± 0.08vs0.30 ± 0.03, P (0.05 or P <0.01). Immunocytochemistry and immunofluorescence results showed that B7-H1 protein was mainly expressed on the cell membrane of 786-0 cells. The expression of B7-H1 in 786-0 cells of IL-2, IFN-α and IFN-γ group was significantly higher than that of 786-0 cells The levels of IL-2 and IFN-α in 786-0 cells were significantly higher than those in 786-0 cells (78.5% ± 0.4%, P <0.01) And IFN-γ upregulated the expression of B7-H1mRNA and protein in 786-0 cells, among which IFN-γ up-regulated and IFN-α up-regulated.