沉默ADAM17抑制肺癌细胞sMICA分泌促进NK细胞的抗肿瘤免疫效应  

Promoting Antitumor Immune Effect of NK Cells by Inhibiting sMICA Secretion of Lung Cancer Cells with Silencing ADAM17

作　　者：彭非 卢经伟 李勇 张艳霞 PENG Fei;LU Jingwei;LI Yong(Jingzhou Chest Hospital, Hubei Jingzhou 434020, China)

机构地区：[1]湖北省荆州市胸科医院呼吸病临床中心,湖北荆州434020

出　　处：《河北医学》2021年第4期529-534,共6页Hebei Medicine

基　　金：湖北省卫生健康委科研立项项目,(编号:WJ2019Q022)。

摘　　要：目的:探讨沉默解聚素-金属蛋白酶17(ADAM17)抑制肺癌细胞可溶性主要组织相容性复合体Ⅰ类相关分子A(sMICA)的分泌对自然杀伤(NK)细胞的抗肿瘤免疫效应的影响。方法:免疫组织化学法检测肺癌组织和癌旁组织中ADAM17的表达,实时定量聚合酶链反应法(qRT-PCR)检测肺癌细胞A549与人正常肺上皮细胞BEAS-2B中ADAM17的mRNA表达。小干扰RNA(siRNA)内源性敲低ADAM17,qRT-PCR检测ADAM17与主要组织相容性复合体Ⅰ类相关分子A(MICA)的mRNA表达,蛋白质免疫印迹法(Western blot)检测ADAM17与MICA的蛋白表达,酶联免疫吸附测定法(ELISA)检测A549细胞上清液sMICA的表达,流式细胞术检测MICA的表达,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在肺癌组织及肺癌细胞A549中高表达。siRNA内源性敲低ADAM17,A549细胞上清中sMICA含量降低(P<0.05),A549细胞MICA表达升高(P<0.05),NK细胞对A549细胞的杀伤活性显著增强(P<0.05)。结论:沉默ADAM17促进肺癌细胞MICA的表达,抑制sMICA的脱落,增强NK细胞对肺癌细胞的杀伤敏感性。Objective:To investigate the effect of silenced disintegrin-metalloproteinase 17(ADAM17)on the anti-tumor immune effect of natural killer(NK)cells by inhibiting the secretion of soluble major histocompatibility complex class I related molecule A(sMICA)of lung cancer cells.Methods:Immunohistochemistry was used to detect the expression of ADAM17 in lung cancer tissues and adjacent tissues.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the mRNA expression of ADAM17 in lung cancer cells A549 and human normal lung epithelial cells BEAS-2B.Small interfering RNA(siRNA)endogenously knocks down ADAM17,qRT-PCR detects the mRNA expression of ADAM17 and major histocompatibility complex class I related molecule A(MICA),Western blot detects ADAM17 and MICA The protein expression of NK cells was detected by enzyme-linked immunosorbent assay(ELISA)to detect the expression of sMICA in the supernatant of A549 cells,flow cytometry was used to detect the expression of MICA,and the lactate dehydrogenase release method was used to detect the killing sensitivity of NK cells to lung cancer cells.Results:ADAM17 was highly expressed in lung cancer tissues and lung cancer cell A549.siRNA endogenously knocked down ADAM17,the content of sMICA in the supernatant of A549 cells decreased(P<0.05),the expression of MICA in A549 cells increased(P<0.05),and the killing activity of NK cells on A549 cells was significantly enhanced(P<0.05).Conclusion:Silencing ADAM17 promotes the expression of MICA of lung cancer cells,inhibits the shedding of sMICA,and enhances the killing sensitivity of NK cells to lung cancer cells.

关 键 词：ADAM17 SMICA MICA NK细胞 肺癌 

分 类 号：R73[医药卫生—肿瘤]