一种新微小RNA在BPDE抑制滋养层细胞侵袭迁移中的作用和调控机制  

Role and regulatory mechanism of a novel miRNA in BPDE-suppressed invasion and migration of human trophoblast cells

作　　者：杨阳 张莹 陈维娜 米辰炀 王晓庆 万叔坤 张慧东 YANG Yang;ZHANG Ying;CHEN Wei-na;MI Chen-yang;WANG Xiao-qing;WAN Shu-kun;ZHANG Hui-dong(West China School of Public Health/West China Fourth Hospital,Sichuan University,Chengdu,Sichuan 610041,China;不详)

机构地区：[1]四川大学华西公共卫生学院/华西第四医院,四川成都610041 [2]中山大学附属第八医院

出　　处：《现代预防医学》2023年第6期1105-1110,共6页Modern Preventive Medicine

基　　金：国家自然科学基金面上项目(82073589);深圳市科创委基础研究面上项目(CYJ20220530144403008,JCYJ20210324114814038)。

摘　　要：目的探索新微小RNA miR-hz01在二羟环氧苯并芘(BPDE)抑制滋养层细胞侵袭迁移中的作用和机制,揭示BPDE抑制滋养层细胞侵袭迁移的新机制。方法采用女性永生化滋养层细胞系Swan 71,进行BPDE染毒,并转染miR-hz01抑制剂对其进行敲低,通过细胞侵袭(Transwell)与划痕实验测定滋养层细胞侵袭迁移情况,采用逆转录-实时荧光定量PCR(RT-qPCR)测定miR-hz01含量,免疫印迹实验(Western blot,WB)实验测定细胞中蛋白激酶B(protein kinase B,PKB or AKT)蛋白表达水平。采用t检验比较两组数据之间的差异。结果BPDE染毒的滋养层细胞中,miR-hz01表达水平是对照组的2.98倍,AKT蛋白表达水平是对照组的45.30%,滋养层细胞的侵袭迁移被抑制。BPDE染毒的滋养层细胞中再转染miR-hz01抑制剂(Inhibitor)或其相应阴性对照(Inhibitor-NC or NC),BPDE+Inhibitor组的AKT蛋白水平是BPDE+NC组的2.08倍,且滋养层细胞侵袭迁移功能有所恢复。结论BPDE染毒的滋养层细胞中,miR-hz01高表达,BPDE通过上调miR-hz01抑制侵袭迁移关键蛋白AKT表达,进而抑制滋养层细胞侵袭迁移,揭示了环境有害因素BPDE抑制滋养层细胞侵袭迁移的新机制。Objective To explore the role and regulatory mechanism of a novel miR-hz01 in BPDE-suppressed invasion and migration of human trophoblast cells.Methods Immortalized female trophoblast cell line Swan 71 cells were exposed with BPDE and/or transfected with miR-hz01 inhibitor.The invasion and migration of trophoblast cells were determined with transwell assays.MiR-hz01 expression levels were determined by RT-qPCR.AKT protein levels were analyzed by Western blotting.Differences were determined using LSD t-test between two groups.Results The expression level of miR-hz01 was increased by 2.98-fold in BPDE-exposed human trophoblast cells relative to the control group,the protein level of AKT was reduced to 45.30%relative to the control group,and the invasion and migration of trophoblast cells were also suppressed.The protein level of AKT was increased by 2.08-fold in BPDE-exposed trophoblast cells after transfection of miR-hz01 inhibitor(BPDE+Inhibitor group)relative to that in BPDE+NC group,which rescued the invasion and migration of trophoblast cells suppressed by BPDE exposure.Conclusion A novel miR-hz01 is identified in human trophoblast cells,which is highly expressed in BPDE-exposed human trophoblast cells.BPDE reduces the expression level of AKT,a key protein functions for invasion and migration,by up-regulating miR-hz01 expression levels,thereby suppressing the invasion and migration of trophoblast cells.This work reveal a novel mechanism in which environmental toxicant BPDE suppresses the invasion and migration of human trophoblast cells.

关 键 词：miR-hz01 BPDE 侵袭迁移 AKT 

分 类 号：R714.2[医药卫生—妇产科学]