Transcriptomic alteration after miR-6126 tranfection into HepG2-NTCP

Aim: Hepatitis B virus (HBV) infection is one of the most serious global health problems. Our previous study revealed that an increase in the miR-6126 serum level over one year of pegylated interferon therapy predicted a decrease in hepatitis B surface (HBs) antigens. We aimed to clarify whether miR-6126 downregulated the expression level of sodium taurocholate cotransporting polypeptide (NTCP), a host cell receptor required for HBV entry. Methods: HepG2-NTCP and PXB cells were utilized to evaluate the expression level of NTCP after transfection with miR-6126. The protein expression level of NTCP was evaluated using Western blot analysis and immunostaining. The expression profile of messenger RNAs was evaluated using next-generation sequencing to search for direct targets of miR-6126. Samples were triplicated; miR-6126-transfected group and negative control.研究目的：乙型肝炎病毒（HBV）感染是全球最严重的公共卫生问题之一。本课题组前期研究显示，在为期1年的聚乙二醇化干扰素治疗过程中，血清miR-6126水平升高可预测乙型肝炎表面（HBs）抗原水平下降。本研究旨在明确miR-6126是否可下调牛磺胆酸钠协同转运多肽（NTCP）的表达水平——该蛋白是HBV入侵宿主细胞所必需的受体。实验方法：本研究采用HepG2-NTCP与PXB细胞，检测转染miR-6126后NTCP的表达水平。通过蛋白质印迹法与免疫染色法评估NTCP的蛋白表达水平。利用下一代测序分析信使RNA（mRNA）的表达谱，以筛选miR-6126的直接靶标。实验设置三次生物学重复，分为miR-6126转染组与阴性对照组。