喉癌Hep2细胞凋亡调控中HLA-B的作用机制  

Regulation mechanism of HLA-B on laryngeal carcinoma Hep2 cells apoptosis

作　　者：郭艳[1] 富伟能[2] 钟鸣[1] 孙开来[2] 

机构地区：[1]中国医科大学口腔医学院中心实验室,辽宁沈阳110007 [2]中国医科大学基础医学院医学遗传学教研室,辽宁沈阳110001

出　　处：《中华肿瘤防治杂志》2011年第9期659-662,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：辽宁省自然科学基金(20092110)

摘　　要：目的:探讨人类白细胞抗原B(HLA-B)在喉癌Hep2细胞凋亡调控中的作用机制。方法:免疫共沉淀结合蛋白质印迹技术鉴定HLA-B与S100结合蛋白A8(S100A8)、S100A8与S100A9及HLA-B与S100A9在体外发生相互作用的情况,免疫细胞化学对HLA-B、S100A8和S100A9在Hep2细胞中的表达进行定位,同时验证三者之间可能的关系;进一步应用RNA干涉技术,通过Real-ti mePCR及蛋白质印迹评估相互作用基因的表达情况。结果:Hep2细胞中HLA-B和S100A8蛋白在体外存在相互作用,而S100A8和S100A9并没有以异源二聚体的形式存在。免疫细胞化学结果显示,Hep2细胞中S100A8蛋白主要表达于细胞质中,HLA-B主要定位于细胞质和细胞膜上,而S100A9蛋白主要分布于细胞核中。另外,与转染PBS及无义小分子干扰(si RNA)组相比,RNA干涉HLA-B基因能明显下调S100A8 mRNA和蛋白的表达水平,F值分别为553.024、603.582,P值均为0.000;而RNA干涉S100A8基因对HLA-B mRNA和蛋白表达变化无显著影响,F值分别为1.266、1.087,P值分别为0.348、0.395。结论:HLA-B与S100A8相互作用,部分通过调节S100A8/Bcl-2的表达而激发喉癌Hep2细胞凋亡发生。OBJECTIVE： To explore the regulation mechanism of HLAB on laryngeal carcinoma Hep2 cells apoptosis. METHODS： Coimmunoprecipitation together with Western blot technologies were used to detect the interaction between HLAB and S100A8, S100A8 and S100A9, HLAB and S100A9 in vitro. Meanwhile, immunocytochemistry was ap plied to localize and identify the possible relationship among HLAB, S100A8 and S100A9 in Hep2 cells. Furthermore, re altime PCR and Western blot were applied to evaluate the ex pression level of interaction genes after RNA interference. RE SULTS： HLA-B protein interacted with S100A8 in vitro, which not heterodimered with S100A9 in Hep2 cells. Immunocytochem istry revealed that S100A8 protein was expressed in cytoplasm, and HLAB was localized in cytoplasm and membrane, while S100A9 was mainly distributed in nucleus. Additionally, RNA interference HLAB could significantly downregulate the mRNA and protein expression level of S100A8 gene compared with PBS transfection and nonsense siRNA transfection groups （F values were 553. 024 and 603. 582, and P value was all 0. 000）, while no significant HLAB mRNA and protein expression differences were obtained undergoing S100A8 gene RNA interference （F values were 1. 266 and 1. 087,and P values were 0. 348 and 0. 395）. CONCLUSION： HLAB interacting with S100A8, triggers laryngeal carcinoma Hep2 cells apoptosis partly trough regulating the expression of S100AS/Bcl-2.

关 键 词：喉肿瘤 细胞凋亡 HLA抗原 免疫沉淀法 

分 类 号：R739.65[医药卫生—肿瘤]