mTORC1基因通过下调YAP/TAZ信号通路抑制小儿肝母细胞瘤发展的机制研究  

Study on the mechanism of mTORC1 gene in inhibiting the development of pediatric hepatoblastoma through down-regulation of YAP/TAZ signaling pathway

作　　者：任慧[1] 陈秀珍 张娟 牛会忠[1] 耿建磊[1] 赵天娇 REN Hui;CHEN Xiuzhen;ZHANG Juan;NIU Huizhong;GENG Jianlei;ZHAO Tianjiao(Hebei Children’s Hospital, Shijiazhuang 050000, Hebei,China;Special Care Hospital of Hebei Province, Shijiazhuang 050000, Hebei,China)

机构地区：[1]河北省儿童医院,河北石家庄050000 [2]河北省优抚医院,河北石家庄050000

出　　处：《现代中西医结合杂志》2021年第17期1840-1843,1856,共5页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：2019年度河北省医学科学研究课题计划项目(20190132)。

摘　　要：目的探讨mTORC1基因在YAP/TAZ信号通路参与肝母细胞瘤发生发展中的调节作用。方法将人肝母细胞瘤HepG2细胞随机分为空白试剂组、空白质粒组、pcDNA3.1-YAP/TAZ质粒组、pcDNA3.1-YAP/TAZ质粒+RAPA组,使用XfectTM转染试剂分别转染空白试剂、空白质粒、pcDNA3.1-YAP/TAZ质粒、pcDNA3.1-YAP/TAZ质粒。转染后24 h采用实时荧光定量PCR法检测YAP和TAZ mRNA表达量。转染后12 h,空白试剂组、空白质粒组、pcDNA3.1-YAP/TAZ质粒组采用正常培养基培养,pcDNA3.1-YAP/TAZ质粒+RAPA组给予含100 nmol/L的RAPA培养,培养后24 h、48 h、72 h采用MTT法测定HepG2细胞增殖能力,并采用Transwell小室测定培养24 h后HepG2细胞的迁移和侵袭能力。结果空白试剂组和空白质粒组YAP、TAZ mRNA表达量比较差异均无统计学意义(P均>0.05);pcDNA3.1-YAP/TAZ质粒组和pcDNA3.1-YAP/TAZ质粒+RAPA组YAP、TAZ mRNA表达量均明显高于空白试剂组和空白质粒组(P均<0.05),pcDNA3.1-YAP/TAZ质粒组和pcDNA3.1-YAP/TAZ质粒+RAPA组比较差异均无统计学意义(P均>0.05)。培养24 h、48 h、72 h后,pcDNA3.1-YAP/TAZ质粒组HepG2细胞增殖活性吸光度均明显高于其余组(P均<0.05),且pcDNA3.1-YAP/TAZ质粒组迁移细胞数和侵袭细胞数均明显多于其余组(P均<0.05)。结论激活mTORC1可能是YAP/TAZ信号通路参与肝母细胞瘤发生发展的重要途径,针对mTORC1的靶向治疗可能在肝母细胞瘤治疗中具有重要前景。Objective It is to explore the regulatory role of mTORC1 gene in the YAP/TAZ signaling pathway involved in the occurrence and development of hepatoblastoma.Methods Human hepatoblastoma HepG2 cells were randomly divided into blank reagent group,blank plasmid group,pcDNA3.1-YAP/TAZ plasmid group,pcDNA3.1-YAP/TAZ plasmid+RAPA group,the blank reagent,blank plasmid,pcDNA3.1-YAP/TAZ plasmid,pcDNA3.1-YAP/TAZ plasmid were ransfected by XfectTM transfection reagent.24 h after transfection,real-time fluorescent quantitative PCR method was used to detect the expression of YAP and TAZ mRNA.12 h after transfection,the blank reagent group,blank plasmid group,pcDNA3.1-YAP/TAZ plasmid group were cultured in normal medium,and the pcDNA3.1-YAP/TAZ plasmid+RAPA group was cultured with medium containing 100 nmol/L RAPA,after culture for 24 h,48 h and 72 h,the proliferation ability of HepG2 cells was measured by MTT method,and the migration and invasion ability of HepG2 cells after culture for 24 h was measured by Transwell chamber.Results There was no significant difference in the expression of YAP and TAZ mRNA between the blank reagent group and the blank plasmid group(P>0.05);the expression levels of YAP and TAZ mRNA in the pcDNA3.1-YAP/TAZ plasmid group and the pcDNA3.1-YAP/TAZ plasmid+RAPA group were significantly higher than those in the blank reagent group and the blank plasmid group(both P<0.05),there was no significant difference between the the pcDNA3.1-YAP/TAZ plasmid group and the pcDNA3.1-YAP/TAZ plasmid+RAPA group(all P>0.05).After culturing for 24 h,48 h,and 72 h,the absorbance of HepG2 cell proliferation activity in the pcDNA3.1-YAP/TAZ plasmid group was significantly higher than that of the other groups(all P<0.05),and the number of migrating cells and invading cells in the pcDNA3.1-YAP/TAZ plasmid group was significantly higher than that of the other groups(all P<0.05).Conclusion Activating mTORC1 may be an important way for YAP/TAZ signaling pathway to participate in the occurrence and development of hepatoblastoma

关 键 词：肝母细胞瘤 哺乳动物雷帕霉素靶蛋白 Yes相关蛋白 

分 类 号：R735.7[医药卫生—肿瘤]