Chen Shunjin, Ying Mingang, Lin Xiuan, Zheng Xiong, Liu Chang, Liu Hui
Department of Head and Neck Surgery, The Tumor Hospital of Fujian Province Fuzhou 350009, Fujian, China.
Int J Clin Exp Med. 2015 Oct 15;8(10):18208-12. eCollection 2015.
This study aims to observe the expression of MHC-class I chain related protein A (MICA) in oral squamous carcinoma cell and explore its effects on NK cells.
Normal oral mucosa epithelial cell line NOK and oral squamous carcinoma cell lines OEC-M1, SAS and SCC-25 were used in this study. MICA expression in the cells was detected by western blotting and RT-PCR methods, sMICA was detected by ELISA method. The cells were transfected by pEGFP-MICA and pEGFP-NC respectively using Lipofectamine 2000 kit. The transfected cells were co-cultured with NK92 cells. Killing activity of NK92 cells was detected by LDH release method and NKG2D was detected by Flow cytometry. ADAM10 and ADAM17 were detected by ELISA method.
MICA expression in OEC-M1, SAS and SCC-25 cells were lower than that of NOK cells (P<0.01), sMICA levels in OEC-M1, SAS and SCC-25 cells were higher than that of NOK cells (P<0.01). Over-expression of MICA in SCC-25 cells could significantly increase the killing activity of NK92 cells (P<0.01), up-regulate NKG2D (P<0.01) and decrease ADAM10 and ADAM17 contents (P<0.01).
MICA expressed lowly in oral squamous cell carcinoma cells, over-expression of MICA could significantly increase the killing activity of NK92 cells, which could be related with the regulation of ADAM.
本研究旨在观察口腔鳞状细胞癌细胞中MHC I类链相关蛋白A（MICA）的表达情况，并探讨其对自然杀伤细胞（NK细胞）的影响。
本研究采用正常口腔黏膜上皮细胞系NOK以及口腔鳞状癌细胞系OEC-M1、SAS和SCC-25。通过蛋白质免疫印迹法和逆转录聚合酶链反应（RT-PCR）方法检测细胞中MICA的表达，采用酶联免疫吸附测定（ELISA）法检测可溶性MICA（sMICA）。使用Lipofectamine 2000试剂盒分别将pEGFP-MICA和pEGFP-NC转染至细胞。将转染后的细胞与NK92细胞共培养。采用乳酸脱氢酶（LDH）释放法检测NK92细胞的杀伤活性，通过流式细胞术检测NKG2D。采用ELISA法检测解聚素金属蛋白酶10（ADAM10）和解聚素金属蛋白酶17（ADAM17）。
OEC-M1、SAS和SCC-25细胞中MICA的表达低于NOK细胞（P<0.01），OEC-M1、SAS和SCC-25细胞中的sMICA水平高于NOK细胞（P<0.01）。SCC-25细胞中MICA的过表达可显著提高NK92细胞的杀伤活性（P<0.01），上调NKG2D（P<0.01）并降低ADAM10和ADAM17的含量（P<0.01）。
MICA在口腔鳞状细胞癌细胞中低表达，MICA的过表达可显著提高NK92细胞的杀伤活性，这可能与ADAM的调节有关。
