丙戊酸钠通过MEK/ERK信号通路上调NKG2D配体表达增强NK细胞对A375人黑素瘤细胞的杀伤  

Valproate up-regulates the expression of NKG2DL through the MEK/ERK signaling pathway to enhance the killing effect of NK cells on A375 human melanoma cells

作　　者：李生存[1] 王永[1] 燕华玲[1] LI Shengcun;WANG Yong;YAN Hualing(Department of Dermatology and Venereology,Affiliated Hospital of Qinghai University,Xining 810001,China)

机构地区：[1]青海大学附属医院皮肤性病科,青海西宁810001

出　　处：《细胞与分子免疫学杂志》2022年第1期32-38,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：青海省科学技术厅项目基金(26346201)。

摘　　要：目的探讨丙戊酸钠(VPA)通过丝裂原激活激酶激酶/细胞外信号调节激酶(MEK/ERK)信号通路对自然杀伤细胞2组成员D(NKG2D)配体表达及自然杀伤(NK)细胞杀伤黑素瘤细胞的影响。方法采用1 mmol/L VPA处理对数生长期的A375细胞24 h,Western blot法检测主要组织相容性复合体Ⅰ类链相关分子A(MICA)、主要组织相容性复合体Ⅰ类链相关分子B(MICB)、磷酸化的MEK(p-MEK)、MEK、磷酸化的ERK1/2(p-ERK1/2)与ERK1/2的蛋白表达水平;流式细胞术检测MICA与MICB的表达;乳酸脱氢酶(LDH)释放法检测NK92细胞对A375黑素瘤细胞的杀伤活性;使用MEK/ERK信号通路抑制剂PD98059联合VPA处理A375细胞,Western blot法检测MICA、MICB蛋白表达,流式细胞术检测MICA与MICB的表达;LDH释放法检测NK92细胞对A375黑素瘤细胞的杀伤活性。非肥胖糖尿病/重症联合免疫缺陷(NOD/SCID)小鼠成瘤实验检测黑素瘤体积变化,免疫组织化学染色法检测MICA与MICB的表达。结果与对照组相比,VPA组A375细胞MICA与MICB的表达升高,NK92细胞对A375细胞的杀伤率增加,p-MEK/MEK、p-ERK1/2/ERK1/2比值升高。与VPA组相比,VPA联合PD98059组A375细胞MICA与MICB的表达降低,NK92细胞对A375细胞的杀伤率降低;与NK92细胞组相比,NK92细胞联合VPA组NOD/SCID小鼠肿瘤体积减少,MICA与MICB表达升高;与NK92细胞联合VPA组相比,NK92细胞联合VPA和PD98059组NOD/SCID小鼠肿瘤体积增加,MICA与MICB表达降低。结论VPA上调黑素瘤细胞MICA与MICB表达,增强NK92细胞对黑素瘤的杀伤活性,可能与MEK/ERK信号通路的激活有关。Objective To investigate the effect of sodium valproate(VPA) on the expression of NKG2D ligand and the killing effect of NK cells on melanoma cells through MEK/ERK signaling pathway. Methods In the group with A375 cells in logarithmic phase treated with 1 mmol/L of VPA for 24 hours, the protein expression levels of MICA, MICB, phosphorylated MEK(p-MEK), MEK, phosphorylated ERK1/2(p-ERK1/2), and ERK1/2 were detected by Western blotting, the expressions of MICA and MICB were detected by flow cytometry, and the killing effect of NK92 cells on A375 cells was detected by lactate dehydrogenase(LDH) release assay. In the group with A375 cells treated with the MEK/ERK signaling pathway inhibitor PD98059 combined with VPA, the protein expressions of MICA and MICB were detected by Western blotting, the expressions of MICA and MICB were detected by flow cytometry, and the killing effect of NK92 cells on A375 cells was detected by LDH release assay. The changes of melanoma volume in non-obese diabetic/severe combined immunodeficiency(NOD/SCID) mice were detected by tumor formation assay, and the expression of MICA and MICB was detected by immunohistochemical staining. Results Compared with those in the control group, the expressions of MICA and MICB of A375 cells, the killing effect of NK92 cells on A375 cells, and the ratios of p-MEK/MEK and p-ERK1/2/ERK1/2 were increased in the VPA group. Compared with those in the VPA group, the expressions of MICA and MICB of A375 cells and the killing effect of NK92 cells on A375 cells were decreased in the group of VPA combined with PD98059. Compared with those in the group of NK92 cells, the tumor volume of NOD/SCID mice was reduced, and the expressions of MICA and MICB were increased in the group of NK92 cells with VPA. Compared with those in the group of NK92 cells with VPA, the tumor volume of NOD/SCID mice was increased, and the expressions of MICA and MICB were decreased in the group of NK92 cells with VPA and PD98059. Conclusion VPA up-regulates the expression of MICA and MICB i

关 键 词：丙戊酸钠(VPA) 主要组织相容性复合体Ⅰ类链相关分子A(MICA) MICB NK92细胞 细胞外信号调节激酶(ERK) 

分 类 号：R965[医药卫生—药理学] Q279[医药卫生—药学]