microRNA-588通过调控USP22基因表达对神经母细胞瘤增殖和迁移的影响及机制研究  

microRNA-588 regulates expression of USP22 gene on proliferation and migration of neuroblastoma and its mechanism research

作　　者：许雅丽[1] 李笃妙[1] 吴强[1] 林俊山[1] XU Yali;LI Dumiao;WU Qiang;LIN Junshan(Department of Pediatrics,the First Affiliated Hospital of Fujian Medical University,Taijiang 350005,China)

机构地区：[1]福建医科大学附属第一医院儿外科,台江350005

出　　处：《中国免疫学杂志》2023年第9期1809-1813,共5页Chinese Journal of Immunology

基　　金：福建省自然科学基金项目(2019J01433)。

摘　　要：目的:探究miR-588通过调控泛素特异性蛋白酶22(USP22)表达对神经母细胞瘤(NB)增殖和迁移的影响及机制。方法:通过双荧光素酶报告验证miR-588与USP22的靶向关系。将SK-N-SH细胞分为NC组、miR-588组、miR-588^(+)USP22组和USP22组,通过转染miR-588 mimic和/或USP22质粒过表达其水平。通过qPCR和Western blot检测USP22 mRNA和蛋白水平验证靶向关系。检测各组细胞的增殖、迁移、侵袭能力。通过ELISA检测免疫抑制相关指标肿瘤坏死因子-α(TNF-α)和可溶性白细胞介素-2受体(sIL-2R)水平。结果:miR-588与USP22 mRNA 3’端的非翻译区域(3’-UTR)通过碱基互补配对的方式结合。miR-588组USP22 mRNA和蛋白显著低于NC组(P<0.05),miR-588^(+)USP22组USP22 mRNA和蛋白水平显著高于miR-588组而低于USP22组(P<0.05)。与NC组比较,miR-588组细胞增殖、迁移、侵袭能力显著升高(P<0.05)。USP22作用与miR-588相反(P<0.05),并且miR-588^(+)USP22组增殖、迁移、侵袭能力显著高于miR-588组而低于USP22组(P<0.05)。与NC组比较,miR-588组TNF-α[(8.97±0.90)pg/ml]和sIL-2R[(6.68±0.75)pg/ml]水平显著升高,USP22组TNF-α[(2.04±0.31)pg/ml]和sIL-2R[(1.48±0.19)pg/ml]水平显著降低(P<0.05),miR-588^(+)USP22组TNF-α[(4.16±0.49)pg/ml]和sIL-2R[(3.21±0.41)pg/ml]水平显著低于mi R-588组但高于USP22组(P<0.05)。结论:miR-588可靶向抑制USP22表达诱导TNF-α和sIL-2R分泌,从而缓解免疫抑制,进而抑制NB细胞增殖和转移。Objective:To explore the effects of miR-588 on the proliferation and migration of neuroblastoma(NB)cells by regulating the expression of ubiquitin-specific protease 22(USP22)and its mechanism.Methods:The targeting relationship between miR-588 and USP22 were verified by dual luciferase report.The SK-N-SH cells were divided into NC group,miR-588 group,miR-588+USP22 group and USP22 group.The levels of miR-588 mimic and/or USP22 were overexpressed by transfection of miR-588 mimic and/or USP22 plasmid.The level of USP22 mRNA and protein were detected by qPCR and Western blot respectively.The proliferation,migration and invasion abilities of each group were detected.The levels of tumor necrosis factor-α(TNF-α)and soluble interleukin-2 receptor(sIL-2R)were detected by ELISA.Results:miR-588 could bind to the 3'-untranslated region(3'-UTR)of USP22 mRNA.The USP22 mRNA and protein in the miR-588 group were significantly lower than those in the NC group(P<0.05).The levels of USP22 mRNA and protein in the miR-588+USP22 group were significantly higher than those of the miR-588 group and lower than those of the USP22 group(P<0.05).Compared with the NC group,the proliferation,migration and invasion abilities of the miR-588 group were significantly increased(P<0.05).The effects of USP22 were opposite to miR-588(P<0.05).The proliferation,migration and invasion abilities of miR-588+USP22 group were significantly higher than those of the miR-588 group and lower than those of the USP22 group(P<0.05).Compared with the NC group,the levels of TNF-α[(8.97±0.90)pg/ml]and sIL-2R[(6.68±0.75)pg/ml]in the miR-588 group were significantly increased,while the TNF-α[(2.04±0.31)pg/ml]and sIL-2R[(1.48±0.19)pg/ml]levels in the USP22 group were significantly reduced(P<0.05).The levels of TNF-α[(4.16±0.49)pg/ml]and sIL-2R[(3.21±0.41)pg/ml]in the miR-588+USP22 group were significantly lower than those in the miR-588 group and higher than those in the USP22 group(P<0.05).Conclusion:miR-588 can induce the secretion of TNF-αand sIL-2R by targ

关 键 词：神经母细胞瘤 miR-588 泛素特异性蛋白酶22 免疫抑制 

分 类 号：R739.44[医药卫生—肿瘤]