不同启动子调控HBV X基因的表达及其对细胞凋亡的影响。

StephenW

Eur Rev Med Pharmacol Sci. 2018 Sep;22(18):5906-5913. doi: 10.26355/eurrev_201809_15919.
Expressions of HBV X gene regulated by different promoters and their effects on cell apoptosis.
Chen W1, Huang L, Liu H, Li XM, Xu CM.
Author information

1
    Department of Gastroenterology, Xiangyang No. 1 People's Hospital, Hubei University of Medicine, Xiangyang, China. [email protected].

Abstract
OBJECTIVE:

  To investigate the difference in expression level of hepatitis B virus (HBV) X gene, and to further study the difference of HBV X protein (HBx) at varied expression levels in apoptosis regulation of hepatocellular carcinoma cells. The recombinant plasmid rPX with HBV enhancer 1 (Enh1), X gene promoter, X gene and polyA tail were constructed, respectively.
PATIENTS AND METHODS:

HepG2 cells were transiently transfected with the recombinant plasmids and control vector pGEM®-7Zf (+) by virtue of Fugene®HD; reverse transcription PCR (RT-PCR) and Western blotting were applied to analyze the transcription and expression of HBV X gene as well as the difference in the expression level in multiple groups. The activity of the transfected cells in each group was detected by using the methyl thiazolyl tetrazolium (MTT) method for 6 consecutive days after transfection. A flow cytometer was utilized to measure the cell apoptosis rate.
RESULTS:

The RT-PCR results showed that messenger RNA expression of HBV X gene was detected in all HepG2 cells transfected by different recombinant plasmids, of which the relationships of the expression levels were rCX>rEX1 and rEX2> rPX (p<0.05). Only HepG2/rCX cells in each group of transfected cells showed HBx expression by Western blot. MTT method revealed that there were notable differences between HepG2/rCX, HepG2/rEX1, HepG2/rPX and HepG2/pGEM®-7Zf (+) (p<0.05). The apoptosis rates of HepG2/rCX, HepG2/rEX1 and HepG2/rPX were significantly higher than that of HepG2/pGEM®-7Zf (+) (p<0.05).
CONCLUSIONS:

HBx can promote cell apoptosis. Results of this research also indicate that there is a significant difference in the pro-apoptotic role of HBx when its expression is regulated by different promoters, and such a difference may be a part of the complex pathogenic mechanisms of HBV and hepatocellular carcinoma.

PMID:
    30280771

StephenW

Eur Rev Med Pharmacol Sci。 2018年9月; 22（18）：5906-5913。 doi：10.26355 / eurrev_201809_15919。
不同启动子调控HBV X基因的表达及其对细胞凋亡的影响。
陈文，黄,,刘,,李晓敏，徐CM。
作者信息

1
    湖北医科大学襄阳市第一人民医院消化内科，襄阳[email protected]。

抽象
目的：

  研究乙型肝炎病毒（HBV）X基因表达水平的差异，进一步研究HBV X蛋白（HBx）在肝细胞癌细胞凋亡调控中不同表达水平的差异。构建了具有HBV增强子1（Enh1），X基因启动子，X基因和polyA尾的重组质粒rPX。
患者和方法：

通过Fugene®HD，用重组质粒和对照载体pGEM®-7Zf（+）瞬时转染HepG2细胞;采用逆转录聚合酶链反应（RT-PCR）和Western blotting分析HBV X基因的转录和表达以及多组表达水平的差异。转染后连续6天用甲基噻唑四唑（MTT）法检测各组转染细胞的活性。流式细胞仪用于测量细胞凋亡率。
结果：

RT-PCR结果显示，不同重组质粒转染的HepG2细胞均检测到HBV X基因的信使RNA表达，其表达水平为rCX> rEX1和rEX2> rPX（p <0.05）。每组转染细胞中只有HepG2 / rCX细胞通过Western印迹显示HBx表达。 MTT法显示HepG2 / rCX，HepG2 / rEX1，HepG2 / rPX和HepG2 /pGEM®-7Zf（+）之间存在显着差异（p <0.05）。 HepG2 / rCX，HepG2 / rEX1和HepG2 / rPX的凋亡率显着高于HepG2 /pGEM®-7Zf（+）（p <0.05）。
结论：

HBx可以促进细胞凋亡。该研究的结果还表明，当HBx的表达受不同启动子调节时，HBx的促细胞凋亡作用存在显着差异，并且这种差异可能是HBV和肝细胞癌的复杂致病机制的一部分。

结论：
    30280771