Ren Yan-Xin, Yang Jie, Sun Rui-Mei, Zhang Li-Juan, Zhao Liu-Fang, Li Bao-Zhong, Li Lei, Long Hai-Ting, Sun Qiang-Ming, Huang Yun-Chao, Li Xiao-Jiang
Head and Neck Tumor Research Center, Third Affiliated Hospital of Kunming Medical University, Kunming, China.
Department of Pathology, Third Affiliated Hospital of Kunming Medical University, Kunming, China.
Cytotechnology. 2016 Dec;68(6):2625-2636. doi: 10.1007/s10616-016-9987-9. Epub 2016 Sep 20.
The HLA-I antigen processing machinery (APM) plays a crucial role in the anticancer immune response. The loss of surface expression of HLA-I molecules is particularly important as this enables tumor cells to evade recognition and lysis by cytotoxic T-lymphocytes. Transcriptional control of the APM genes is regulated by the nuclear factor kappa B (NF-κB). BCRFl is an Epstein-Barr virus homologue of human IL-10 (hIL-10) and is known as viral IL-10 (vIL-10). vIL-10 shares many immunosuppressive effects with hIL-10 but lacks the immunostimulatory effect of hIL-10. The aim of this study was to assess whether vIL-10 inhibits APM components (TAP-1, TAP-2, LMP-2, LMP-7 and HLA-I) through the NF-κB signaling pathway in nasopharyngeal carcinoma. This work demonstrated that vIL-10 inhibited NF-κB activation by blocking IKK phosphorylation and promoting the expression of IKB. TNF-α treatment led to a strong translocation of NF-κB p65, whereas pretreatment with vIL-10 before TNF-α treatment blocked NF-κB p65 translocation. vIL-10 also inhibited TNF-α-induced DNA-binding of NF-κB p65 in the nucleus. Furthermore, chromatin immunoprecipitation analysis demonstrated that NF-κB p65 could bind to the TAP-1, TAP-2, LMP-2, LMP-7 and HLA-I gene promoters, and after TNF-α stimulation, the down-regulation of TAP-1, TAP-2, LMP-2, LMP-7 and HLA-I transcription by vIL-10 correlated with the suppression of NF-κB in CNE-2 cells. Surprisingly, vIL-10 inhibits only TAP-1 and LMP-7 transcription in CNE-1 cells. Taken together, these results suggest that the inhibition of NF-κB activity may be an important mechanism for vIL-10 suppression of APM (TAP-1, TAP-2, LMP-2, LMP-7 and HLA-I) gene transcription in CNE-2 cells.
人类白细胞抗原I类分子抗原加工机制(APM)在抗癌免疫反应中发挥着关键作用。HLA-I分子表面表达的缺失尤为重要,因为这使得肿瘤细胞能够逃避细胞毒性T淋巴细胞的识别和裂解。APM基因的转录调控由核因子κB(NF-κB)介导。BCRF1是人类IL-10(hIL-10)的爱泼斯坦-巴尔病毒同源物,被称为病毒IL-10(vIL-10)。vIL-10与hIL-10具有许多免疫抑制作用,但缺乏hIL-10的免疫刺激作用。本研究的目的是评估vIL-10是否通过NF-κB信号通路抑制鼻咽癌中APM成分(TAP-1、TAP-2、LMP-2、LMP-7和HLA-I)。这项工作表明,vIL-10通过阻断IKK磷酸化和促进IκB的表达来抑制NF-κB的激活。TNF-α处理导致NF-κB p65强烈易位,而在TNF-α处理前用vIL-10预处理可阻断NF-κB p65易位。vIL-10还抑制TNF-α诱导的NF-κB p65在细胞核中的DNA结合。此外,染色质免疫沉淀分析表明,NF-κB p65可与TAP-1、TAP-2、LMP-2、LMP-7和HLA-I基因启动子结合,在TNF-α刺激后,vIL-10对TAP-1、TAP-2、LMP-2、LMP-7和HLA-I转录的下调与CNE-2细胞中NF-κB的抑制相关。令人惊讶的是,vIL-10仅抑制CNE-1细胞中TAP-1和LMP-7的转录。综上所述,这些结果表明,抑制NF-κB活性可能是vIL-10抑制CNE-2细胞中APM(TAP-1、TAP-2、LMP-2、LMP-7和HLA-I)基因转录的重要机制。