缺氧微环境下ADAM17与ERp5的高表达对肺癌细胞逃避NK细胞免疫杀伤的机制研究  

Effects of the upregulated ADAM17 and ERp5 on lung cancer cells evasion from NK cell immune killing under hypoxic microenvironment

作　　者：张嫩英 余瑞云 徐震[2] ZHANG Nenying;YU Ruiyun;XU Zhen(Department of General Practice,Xinxiang Central Hospital,Xinxiang 453000,China;Department of Respiratory and Critical Care Medicine,Xinxiang Central Hospital,Xinxiang 453000,China)

机构地区：[1]新乡市中心医院全科医学科,453000 [2]新乡市中心医院呼吸及危重症医学科一,453000

出　　处：《免疫学杂志》2021年第4期308-315,共8页Immunological Journal

摘　　要：目的探讨缺氧条件下ADAM17与ERp5的高表达对肺癌细胞逃避NK细胞免疫杀伤的影响。方法免疫组织化学法检测肺癌组织和癌旁组织中ADAM17与ERp5的表达;qRT-PCR与Western blot检测缺氧条件下的肺癌细胞中ADAM17、ERp5、MICA与MICB的表达;小干扰RNA内源性敲低ADAM17与ERp5;酶联免疫吸附测定法检测缺氧条件下的肺癌细胞上清中sMICA与sMICB的含量;流式细胞术检测CD3^(-)CD56^(+)NK细胞的纯度;乳酸脱氢酶释放法检测NK细胞对肺癌细胞的杀伤敏感性。结果ADAM17与ERp5在肺癌组织中高表达;缺氧条件下,肺癌细胞中ADAM17与ERp5的表达量升高(P<0.05);CD3^(-)CD56^(+)NK细胞的纯度高于90%;缺氧条件下,与si-NC组相比,si-ADAM17与si-ERp5组肺癌细胞上清中sMICA与sMICB的含量降低(P<0.05),肺癌细胞中MICA与MICB的表达升高(P<0.05),NK细胞对肺癌细胞的杀伤活性增强(P<0.05);与siADAM17与si-ERp5组相比,si-ADAM17+si-ERp5组肺癌细胞上清中sMICA与sMICB的含量降低(P<0.05),肺癌细胞MICA与MICB的表达升高(P<0.05),NK细胞对肺癌细胞的杀伤活性增强(P<0.05)。结论缺氧条件下,沉默ADAM17与ERp5通过抑制肺癌细胞中sMICA与s MICB的分泌,促进MICA与MICB的表达,增强了NK细胞对肺癌细胞的免疫杀伤。This study was designed to explore the effect of the up regulated ADAM17/ERp5 on lung cancer cells escaping from NK cell immune killing under hypoxic condition.Immunohistochemistry was performed to detect the expression of ADAM17 and ERp5 in lung cancer tissues and adjacent tissues.qRT-PCR and Western blot were used to detect the expression of ADAM17,ERp5,MICA and MIAB in lung cancer cells under hypoxic condition.Small interfering RNA was transfected into the lung cancer cells to endogenously suppress ADAM 17 and ERp5.Enzyme linked immunosorbent assay was employed to detect the content of sMICA and sMICB in the supernatant of lung cancer cells under hypoxia,while flow cytometry was used to detect the purity of CD3^(-)CD56^(+)NK cells.Lactate dehydrogenase release method was used to detect cytotoxicity of NK cells against lung cancer cells.Data showed that ADAM17 and ERp5 were highly expressed in lung cancer tissues,and the expression of ADAM17 and ERp5 in lung cancer cells were increased under the hypoxic condition.The purity of CD3-CD56’NK cells was higher than 90%.Compared with the si-NC group,the content of sMICA and sMICB in the supernatant of lung cancer cells were decreased,the expression of MICA and MICB were increased in lung cancer cells,and the cytotoxicity of NK cells against lung cancer cells was enhanced under hypoxic conditions in si-ADAM17 and si-ERp5 groups.Compared with si-ADAM17 and si-ERp5 groups,the content of sMICA and sMICB in the supernatant of lung cancer cells was decreased,and the cytotoxicity of NK cells against lung cancer cells was enhanced under hypoxic condition in the si-ADAM17+si-ERp5 group.Taken together,Under hypoxic conditions,silencing ADAM17 and ERp5 could inhibit the secretion of sMICA and sMICB,promote the expression of MIGA and MIGB in lung cancer cells,and enhance the immune killing of NK cells against lung cancer cells.

关 键 词：缺氧微环境 ADAM17 ERp5 MICA MICB NK细胞 肺癌 

分 类 号：R734.2[医药卫生—肿瘤]