【摘要】 目的用实时荧光定量聚合酶链反应(FQ-PCR)方法检测不同亚型干扰素α(IFN-α2b、IFN-α2a、IFN-α1b)对HepG2细胞内信号传导分子STAT1mRNA表达的影响。方法1000IU/mlIFN-α2b、IFN-α2a、IFN-α1b分别作用于HepG2细胞4、8、16、24h后,用RT-PCR的方法扩增STAT1目的片段,用T-A克隆方法构建定量的标准模板,并用荧光定量PCR方法观察IFN-α2b、IFN-α2a、IFN-α1b作用后HepG2细胞内STAT1mRNA的表达水平。结果经IFN-α2b、IFN-α2a、IFN-α1b处理后,HepG2细胞内的STAT1mRNA水平较未用IFN-α的空白对照组明显上调,并于IFN-α处理8h后STAT1mRNA水平达高峰。此时,IFN-α2b、IFN-α2a和IFN-α1b的STAT1mRNA水平(×107copies/ml)依次为3·59±0·25、2·73±0·43和4·85±0·55,三者之间存在显著性差异。结论IFN-α作用于HepG2细胞后,IFN-α1bSTAT1mRNA的表达水平最高,IFN-α2b次之,IFN-α2a最弱。间接说明IFN-α1b的抗HBV活性较强,IFN-α2b次之,IFN-α2a较弱。更多还原

【Abstract】 Objective To detect the effects of interferon-alpha subtypes(IFN-α 2b,IFN-α 2a and IFN-α 1b)on signaling transduction molecule STAT1mRNA expression in HepG2 cells by real-time fluorescence quatitive PCR(FQ-PCR).Methods After HepG2 cells were treated with 1000IU/ml IFN-α 2b,IFN-α 2a or IFN-α 1b for 4,8,16,24h, respectively,we constructed the quantitative standard template with T-A clone methods using the conventional RT-PCR to amplify STAT1 gene from cultured HepG2 cells.STAT1 mRNA expression after pre-treatment with IFN-α 2b,IFN-α 2a or IFN-α 1b was determined with FQ-PCR.Results HepG2 cells possessed relatively low basal mRNA levels of STAT1,and their expression was greatly up-regulated by IFN-α.STAT1 mRNA level reached the maximum after treatment with IFN-α,at which,STAT1 mRNA levels(×10~7copies/ml) were 3.59±0.25,2.73±0.43 and 4.85±0.55 after treated with IFN-α 2b,IFN-α 2a or IFN-α 1b, respectively. It showed statistical difference among the three groups.Conclusion STAT1mRNA expression level was in an order of IFN-α 1b>IFN-α 2b>IFN-α 2a aftre pre-treatmented with IFN-α,suggesting that the antiviral effect of IFN-α 1b is stronger than that of IFN-α 2b and IFN-α 2a.更多还原