摘要目的 观察乙型肝炎病毒编码X蛋白(HBX)对肝脏卵圆细胞中转化生长因子-β(TGF-β)/Smad中核心元件Smad2、Smad3、Smad4表达的调控作用。方法 稳定转染HBX基因的大鼠卵圆细胞株HBX-EGFP-LE/6作为实验组,转染绿色荧光蛋白标记空载体的大鼠卵圆细胞株EGFP-LE/6,大鼠卵圆细胞株LE/6作为对照组。Real-time聚合酶链反应(PCR)与Western blot分别检测3种细胞株中Smad2、Smad3、Smad4的mRNA和蛋白表达。结果 对照组Smad3的mRNA表达量分别是实验组的(4.56±0.79)倍和(4.14±0.38)倍,差异有统计学意义(P＜0.01)。对照组Smad4的mRNA表达量分别是实验组的(1.41±0.04)倍和(1.63±0.43)倍,差异有统计学意义(P＜0.01)。实验组Smad3蛋白表达水平有一定程度降低。结论 HBX可以在卵圆细胞中影响TGF-β/Smad信号通路中的核心元件Smad3的转录及蛋白合成,对Smad4的转录有一定影响。

abstractsObjective To investigate the effect of X protein of hepatitis B virus (HBX) on the expression of Smad2, Smad3 and Smad4. Methods Intracellular Smad2, Smad3 and Smad4 expression was detected by real-time polymerase chain reaction (PCR) in HBX-transfected rat oval cells named HBX-EGFP-LE/6, and LE6, EGFP-LE/6 control oval cells. The protein levels of Smad2, Smad3 and Smad4 were determined by Western blotting in oval cells. Results The Smad 3 mRNA and protein expression levels in LE/6 and EGFP-LE/6 cells were (4. 56 ±0. 79) and (4. 14 ±0. 38) folds of those in HBX-EGFP-LE/6 cells (P ＜ 0. 01 ). Smad3 protein levels were down-regulated and the mRNA levels of Smad4 in LE/6 and EGFP-LE/6 cells were ( 1.41 ± 0. 04) and ( 1.63 ± 0. 43 ) folds of those in HBX-EGFP-LE/6 cells ( P ＜0. 01 ). Conclusion HBX expression in rat LE/6 oval cells was sufficient to modulate the mRNA and protein expression of Smad3 in rat oval cells, and mRNA expression of Smad4 was also slightly changed by HBX.