Schlafen基因家族对HBV复制的影响

Effects of Schlafen gene family on HBV replication

目的研究干扰素(interferon,IFN)刺激基因Schlafen(SLFN)对乙型肝炎病毒(hepatitis B virus,HBV)复制的调控作用。方法首先用不同剂量的IFN-α处理肝癌细胞HepG248 h,或在同一剂量下处理不同时间,通过荧光定量PCR(q-PCR)检测SLFN家族基因转录的表达水平;接着在TCGA数据库采用t检验分析HBV感染的肝癌组织与癌旁组织中SLFN基因家族表达差异;进一步通过siRNA干扰和过表达技术,结合q-PCR、Western blot与Southern blot分析病毒复制水平的变化。结果在SLFN基因家族中,只有SLFN5受IFN-α诱导,且呈浓度梯度依赖性;TCGA数据库分析表明在SLFN基因家族中,SLFN5和SLFN11在HBV感染的肝癌组织和癌旁组织中表达明显有差异;在HepG2肝癌细胞中,siRNA干扰SLFN5,HBV复制水平无明显变化;而在Huh7肝癌细胞中,干扰SLFN11后,HBV复制水平增加了1.79倍,而病毒HBc蛋白没有明显变化;在HepG2细胞中过表达SLFN11后,HBV复制中间体水平下降34.67%。结论在肝癌细胞HepG2中,SLFN5受IFN-α诱导,但缺少调控HBV复制的作用;而SLFN11的表达与HBV复制相关,能抑制HBV核心颗粒DNA复制。

Objective To investigate the regulatory effects of interferon-stimulated gene Schlafen(SLFN)on hepatitis B virus(HBV)replication.Methods Firstly,HepG2 cells were treated with IFN-αat different concentrations for 48 h or the same concentration for different periods of time.Expression of SLFN family genes was detected by quantitative real-time PCR(q-PCR).Secondly,the expression of SLFN gene family at mRNA level in HBV-infected tumor tissues and non-tumor tissues recorded in The Cancer Genome Atlas(TCGA)database was compared using t test.Furthermore,changes in the HBV DNA levels after the interference of small interfering RNA(siRNA)-mediated knockdown and overexpression of SLFN5 and SLFN11 genes were analyzed by q-PCR,Western blot and Southern blot.Results Among the SLFN gene family,only SLFN5 expression was induced by IFN-αin a concentration-dependent manner.TCGA database analysis showed that the expression of both SLFN5 and SLFN11 in HBV-infected tumor tissues and non-tumor tissues was significantly different.In HepG2 cells,knocking down the expression of SLFN5 had no obvious effect on the levels of intracellular HBV DNA.It was observed that the level of intracellular HBV DNA increased 1.79 folds in Huh7 cells after knockdown of SLFN11 expression,while no significant change in HBc protein was detected.Conversely,overexpression of SLFN11 induced a 34.67%decrease in intracellular HBV DNA in HepG2 cells.Conclusions In HepG2 cells,SLFN5 expression was induced by IFN-α,but had no effect on HBV replication.However,SLFN11 could inhibit the HBV DNA replication in nucleocapsid.