不同亚型IFN-α对STAT1表达的影响  

The effects of interferon-alpha subtypes on STAT1mRNA expression

作　　者：卢年芳[1] 郑瑞强[1] 林华[1] 黄爱龙[2] 杨德刚[1] 

机构地区：[1]扬州大学医学院附属医院,225001 [2]重庆医科大学病毒性肝炎研究所

出　　处：《江苏医药》2006年第10期901-903,共3页Jiangsu Medical Journal

基　　金：国家自然科学基金项目(30300298)

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。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 realtime 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 1 b 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 rnRNA 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 1FN-α 2b,IFN-α 2a or IFN-α 1b, respectively. It showed statistical difference among the three groups. Conclusion STATlmRNA expression level was in an order of IFN-α 1b〉IFN-α 2b^IFN-a 2a aftre pre-treatmented with IFN-a, suggesting that the antiviral effect of IFN-α 1b is stronger than that of IFN-a 2b and IFN-α 2a.

关 键 词：干扰素-Α 荧光定量聚合酶链反应 基因重组 

分 类 号：R512.63[医药卫生—内科学]