| 孟诗敏,陈忠伟,李涵泺,魏艳红,胡康洪.半乳糖凝集素-3结合蛋白作为正向调节因子参与IFN-γ介导的抗HBV作用[J].现代生物医学进展英文版,2021,(23):4441-4449. |
| 半乳糖凝集素-3结合蛋白作为正向调节因子参与IFN-γ介导的抗HBV作用 |
| LGALS3BP Acts as a Positive Regulator and is Involved in IFN-γ Mediated Anti-HBV Process |
| Received:March 15, 2021 Revised:April 13, 2021 |
| DOI：10.13241/j.cnki.pmb.2021.23.009 |
| 中文关键词: 乙型肝炎病毒 干扰素γ 半乳糖凝集素-3结合蛋白 增殖 |
| 英文关键词: Hepatitis B virus (HBV) Interferon gamma (IFN-γ) Galactose lectin-3 binding protein (LGALS3BP) Proliferation |
| 基金项目:湖北工业大学高层次人才启动基金项目（337203） |
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| 中文摘要: |
| 摘要 目的：研究半乳糖凝集素-3结合蛋白（LGALS3BP）在IFN-?酌介导的抗HBV中的作用。方法：采用qRT-PCR及Western Blot分别检测HepG2细胞瞬转过表达质粒及干扰质粒后LGALS3BP转录水平和翻译水平的变化及检测IFN-γ对HepG2细胞内源性LGALS3BP转录水平和翻译水平的影响；采用CCK-8评价LGALS3BP表达量的变化对HepG2细胞增殖的影响及确定IFN-γ处理HepG2和HepG2.215的最适浓度；采用ELISA检测当LGALSEBP表达量发生变化时，IFN-γ刺激后对细胞外分泌的HBsAg和HBeAg的影响；采用q-PCR和RT-PCR分别检测细胞内HBV核衣壳中DNA和细胞内HBV RNA水平。结果：40 ng/mL的IFN-γ处理HepG2细胞48小时，LGALS3BP mRNA表达量与对照组相比提高2.87倍，但LGALS3BP表达量的变化对细胞增殖无显著作用；当过表达LGALS3BP时，在IFN-γ介导的抗HBV过程中，与对照组相比，HepG2.215细胞分泌的HBsAg、HBeAg以及细胞内HBV DNA和RNA的相对下降幅度分别为46%、67%、59%、49%；与对照组相比，转染pCH9-3091的HepG2细胞分泌的HBsAg、HBeAg以及细胞内HBV DNA和RNA的相对下降幅度分别为67%、53%、60%、29%；而当干扰LGALS3BP时，在IFN-γ介导的抗HBV过程中，与对照组相比，HepG2.215细胞分泌的HBsAg、HBeAg以及细胞内HBV DNA和RNA的相对上升幅度分别为46%、67%、59%、49%；与对照组相比，转染pCH9-3091的HepG2细胞分泌的HBsAg、HBeAg以及细胞内HBV DNA和RNA的相对上升幅度分别为67%、77%、67%、45%。结论：LGALS3BP作为正向调节因子参与IFN-?酌介导的抗HBV过程。 |
| 英文摘要: |
| ABSTRACT Objective: This study was to investigate the role of LGALS3BP in the IFN-γ mediated anti-HBV process in human hepatocytes in vitro. Methods: HepG2 and HepG2.215 cell lines were transiently transfected with LGALS3BP overexpression plasmid and its interfering plasmid, and levels of LGALS3BP transcription and translation were detected by means of qRT-PCR and Western Blot. CCK-8 was used to determine the optimal concentration of IFN-γ treatment for HepG2 and HepG2.215 cells, and evaluate the cell proliferation. The effects of IFN-γ treatment on transcription and translation of endogenous LGALS3BP in HepG2 cells were investigated using qRT-PCR and Western Blot. When changing LGALS3BP expression, the effect of IFN-γ stimulation on the exocrine HBsAg and HBeAg were examined using ELISA, and levels of DNA in HBV nucleocapsid and intracellular RNA were evaluated using q-PCR and RT-PCR, respectively. Results: After treated with 40 ng/ml IFN-γ for 48 hours, the level of LGALS3BP mRNA in HepG2 cells was 2.87 folds higher than that in the control group, whereas the change of LGALS3BP expression had no significant effect on the cell proliferation. In case that LGALS3BP was overexpressed in HepG2.215 cells during the IFN-γ mediated anti-HBV process, compared with the control group, it was found the relative decrease rates of extracellular HBsAg, HBeAg as well as of the relative decrease rates of intracellular HBV DNA- and RNA levels were 46%, 67%, 59% and 49%, respectively; For HepG2 cells that were transiently transfected with pCH9-3091, compared with the control group, the relative decrease rates of extracellular HBsAg, HBeAg as well as the intracellular viral DNA and RNA were 67%, 53%, 60% and 29%, respectively. In contrast, in case that LGALS3BP was down-regulated by siRNA interfering-plasmid SHLG, in the process of IFN-γ mediated anti HBV for HepG2.215, compared with the control group, the relative increase rates of secreted HBsAg, HBeAg as well as of intracellular viral DNA- and RNA levels were 46%, 67%, 59% and 49%, respectively; For HepG2 that transiently transfected with pCH9-3091, the relative increase rates of HBsAg, HBeAg as well as of intracellular viral DNA and RNA levels were 67%, 77%, 67% and 45%, respectively. Conclusion: LGALS3BP is a positive regulator and participates in the IFN-γ mediated anti-HBV process. |
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