拉米夫定抑制乙型肝炎病毒X基因的转录与表达

Inhibition of transcription and expression of X gene by lamivudine

目的 乙型肝炎病毒(HBV)X蛋白在HBV相关性肝癌的发生中有着重要作用,本研究旨在探讨拉米夫定对HBV X基因复制、转录及表达水平的影响。 方法 用聚合酶链反应法检测拉米夫定对转染X基因复制的影响,用逆转录-聚合酶链反应法检测拉米夫定对转染X基因mRNA的影响,用生物分子相互作用系统检测拉米夫定对X蛋白表达的影响,探讨了拉米夫定作用的量效和时效关系。 结果 拉米夫定组与对照组HBV X基因复制的目的基因条带吸光度(A)值分别是151.4±3.5和144.0±11.4,差异无统计学意义。4.36×104mol/L拉米夫定持续作用24 h能完全抑制X基因的转录,对照组与治疗组(16 h)目的条带的A值分别为243.9±9.0和133.2±7.8,P<0.01,其抑制效应随药物浓度增加和作用时间延长而增强。4.36×104mol/L拉米夫定持续作用16 h,使代表X蛋白表达的最大结合量值从353.3±15.9降至252.3±18.8,P<0.01。结论 拉米夫定不影响HepG2x中转染的X基因复制,但对X基因的转录和蛋白表达有明显抑制作用。

Objective The X protein of the hepatitis virus B plays an important role in the development of hepatocellular carcinoma (HCC). In this experiment we studied the effects of lamivudine on replication, transcription and expression of the X gene. Methods The replication of transfected X gene was measured by polymerase chain reaction. The mRNA of transfected X gene was analyzed by reverse transcriptase polymerase chain reaction. The expression of X protein was detected by biosensor. Moreover, we studied if these effects changed in a dose and time dependent manner. Results The OD data of purpose band reflected the replication of X gene for control and lamivudine groups were 151.4 ± 3.5 and 144.0 ± 11.4, respectively, P > 0.05. The transcription of mRNA for X gene could be inhibited by lamivudine at a concentration of 4.36 × 10-4 mol/L for 24 h. The OD data of purpose band for control and treatment groups (16 h) were 243.9 ± 9.0 and 133.2 ± 7.8, P < 0.01. The inhibition was enhanced with the increase of lamivudine concentration and reacting time. The expression of the X protein in HepG2x cells was suppressed by lamivudine at a concentration of 4.36 × 10-4 mol/L for 16 hours. The resonance unit for expression of the X protein decreased from 353.3 ± 15.9 to 252.3 ± 18.8, P < 0.01. Conclusion Although the replication of transfected X gene of HepG2x cells is not influenced by lamivudine, it may significantly inhibit the transcription and expression of the X protein.