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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 investigate the effects of IL-2, IFN-α and IFN-γ on the expression of B7-H4 in human clear cell carcinoma 786-0 cells. Methods: The expression of B7-H4 mRNA in 786-0 cells was detected by RT-PCR after treatment with IL-2, IFN-α and IFN-γ. The expression of B7-H4 mRNA was detected by ELISA and immunocytochemistry Protein expression. Results: The results of RT-PCR showed that the expression of B7-H4 mRNA was significantly higher in IL-2 group (0.75 ± 0.06), IFN-α group (0.68 ± 0.05) and IFN-γ group (0.95 ± 0.08) Higher than untreated cells (0.30 ± 0.03) (P <0.05). The results of immunocytochemical staining showed that the expression of B7-H4 protein was detected in the membrane and cytoplasm of 786-0 cells. The expression of B7-H4 protein in 786-0 cells was increased by IL-2, IFN-α and IFN-γ. The results of ELISA showed that IFN-|Ã (44.89 +/- 0.97) ng / ml and IFN-|Ã group [(46.74 +/- 2.25) ng / ml and IFN- The expression of secreted B7-H4 in 0 cell supernatant was significantly higher than that in untreated group [(34.42 ± 1.69) ng / ml] (P <0.05). Flow cytometry results showed that IL-2 [(44.89 ± 0.94)%], IFN-α [46.41 ± 0.55]% and IFN-γ The positive expression rate of B7-H4 protein on the surface was significantly higher than that of the untreated group [(30.45 ± 0.96)%] (P <0.05). CONCLUSION: IL-2, IFN-α and IFN-γ up-regulate the expression of B7-H4 in 786-0 cells both in transcription and translation, of which IFN-γ up-regulates the strongest.