摘要
慢性乙肝病毒感染是亚洲国家肝癌的常见病因,乙肝病毒所表达的乙肝病毒X蛋白(Hepatitis B virus X protein,HBx)是肝癌发生发展的重要推动因子。核因子-κB(Nuclear factorκB,NF-κB)是模式识别受体下游重要的转录因子,肝细胞生长因子(Hepatocyte growth factor,HGF)/c-Met途径能够促进NF-κB活化,并通过活化的NF-κB来调节肝癌细胞的迁移、侵袭等生物学环节。但HBx是否直接通过NF-κB通路来调节肝癌细胞的迁移、侵袭尚未明确。为探究HBx通过NF-κB通路调节肝癌细胞迁移、侵袭的作用,本研究采用培养肝癌HepG2细胞并分组,空白对照组用不含药物及质粒的DMEM处理,空白质粒组转染1.2μg的pcDNA3.1空白质粒,HBx质粒组转染不同浓度的HBx表达质粒pcDNA3.1-HBx,HBx+PDTC组转染1.2μg的HBx表达质粒pcDNA3.1-HBx并用含有50μmol/L PDTC的DMEM进行处理;皮下注射HepG2细胞建立移植瘤小鼠模型,称量移植瘤的质量。检测细胞迁移及侵袭活力、细胞及移植瘤中NF-κB通路分子、迁移基因、侵袭基因的表达。结果显示,与空白对照组、空白质粒组比较,HBx质粒组细胞中NF-κB、HGF、c-Met、N-cadherin、Vimentin、MMP2、MMP9的蛋白表达水平及相对愈合面积、侵袭数目均明显增多(P<0.05);与HBx质粒组比较,HBx+PDTC组细胞中NF-κB、HGF、c-Met、N-cadherin、Vimentin、MMP2、MMP9的蛋白表达水平及相对愈合面积、侵袭数目均明显减少(P<0.05);与空白对照组、空白质粒组比较,HBx质粒组移植瘤的质量及移植瘤中NF-κB、HGF、c-Met的蛋白表达水平明显增加(P<0.05)。本研究得出结论,HBx能够促进肝癌细胞的迁移、侵袭,且该作用与激活NF-κB通路有关。
Chronic infection with the hepatitis B virus(HBV)is a common cause of hepatocellular carcinoma(HCC)in Asian countries. Hepatitis B virus X protein(HBx)expressed by hepatitis B virus is an important driving factor for the occurrence and development of HCC. Nuclear factor κB(NF-κB) is an important transcription factor downstream of pattern recognition receptors. The hepatocyte growth factor(HGF)/c-Met pathway can promote NF-κB activation and regulate the migration and invasion of HCC cells by activating NF-κB. However,whether HBx regulates the migration and invasion of HCC cells through the NF-κB pathway directly is not known. We investigated the role of HBx in regulating the migration and invasion of HCC cells through the NF-κB pathway. HepG2 cells were cultured and divided into groups. The blank control group was treated with Dulbecco’s modified Eagle’s medium(DMEM)without drugs or plasmids. The blank plasmid group was transfected with 1.2 μg of the pcDNA3.1 blank plasmid. The HBx plasmid group was transfected with 1.2 μg of the HBx expression plasmid pcDNA3.1-HBx. The HBx+PDTC group was transfected with 1.2μg of the HBx expression plasmid pcDNA3.1-HBx and treated with DMEM containing 50 μmol/L PDTC. A transplanted tumor model in mice was established by subcutaneous injection of HepG2 cells,the weight of the transplanted tumor was documented. Cell migration,invasive activity,migratory genes,invasive genes and expression of molecules in the NF-κB pathway were detected. Results showed that,compared with the blank control group and blank plasmid group,expression of NF-κB,HGF,c-Met,N-cadherin,vimentin,matrix metalloproteinase(MMP)2,MMP9,the relative healing area,and the number of invading HepG2 cells in HBx plasmid group were increased significantly(P<0.05). Compared with the HBx plasmid group,expression of NF-κB,HGF,c-Met,N-cadherin,vimentin,MMP2,MMP9,the relative healing area and the number of invading HepG2 cells in the HBx+PDTC group were decreased significantly(P<0.05). Compared with the blank control group and blank plasmid group,the weight of transplanted tumors and expression of NF-κB,HGF,and c-Met in transplanted tumors were increased significantly(P<0.05). These data suggest that HBx promotes the migration and invasion of HCC cells,and that this effect is related to activation of the NF-κB pathway.
作者
钟细涛
宓余强
ZHONG Xitao;MI Yuqiang(Tianjin Medical University,Tianjin 300070,China;Tianjin Second People's Hospital,Tianjin 300192,China)
出处
《病毒学报》
CAS
CSCD
北大核心
2020年第3期407-414,共8页
Chinese Journal of Virology
关键词
乙肝病毒X蛋白(HBx)
肝癌细胞
迁移
侵袭
Hepatitis B virus X protein(HBx)
Hepatocellular carcinoma cells
Migration
Invasion
