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HBX在肝癌细胞中通过下调miR-199a增强AKT的表达

, PP. 996-1000

Keywords: 乙型肝炎病毒X蛋白,微小RNA,肝癌,AKT信号通路

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Abstract:

目的探讨乙型肝炎病毒X蛋白(hepatitisBvirusXprotein,HBX)调节miR-199a的表达在HBV相关性肝癌发生中的分子机制。方法重组HBX腺病毒(Ad-HBX)感染人肝癌HepG2细胞,HBX的siRNA转染稳定表达HBV基因的人肝癌HepG2.2.15细胞,荧光定量PCR方法检测HBV相关性肝癌组织标本和肝癌细胞中HBX、miR-199a、AKT的mRNA表达水平及运用Westernblot检测肝癌组织标本和肝癌细胞中HBX、AKT的蛋白表达水平;分别用miR-199amimics和miR-199ainhibitor的转染肝癌细胞后,荧光定量PCR方法检测肝癌细胞中AKT的mRNA表达水平及运用Westernblot检测肝癌细胞中AKT的蛋白表达水平。结果miR-199a在HepG2.2.15细胞中表达水平显著低于HepG2细胞,并且证实其表达下调受到了HBX的调控(P<0.05),HepG2.2.15细胞中AKT表达水平显著高于HepG2细胞(P<0.05),在HepG2.2.15细胞中过表达miR-199a能明显下调AKT表达水平以及在HepG2细胞中抑制miR-199a能明显上调的AKT表达水平(P<0.05),此外,miR-199a的表达量在HBV相关性肝癌组织比相应的癌旁组织表达降低。结论HBX可以通过miR-199a调节AKT导致HBV相关性肝癌发生。

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