论文部分内容阅读
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 Hepatitis B virus (HBV) X protein plays an important role in the development of HBV-related hepatocellular carcinoma. This study aimed to investigate the effects of lamivudine on HBV X gene replication, transcription and expression. Methods The effect of lamivudine on the replication of transfectant X gene was detected by polymerase chain reaction. The effect of lamivudine on the mRNA expression of X gene was detected by reverse transcriptase-polymerase chain reaction. The biomolecular interaction system To detect the impact of lamivudine on the expression of X protein, explore the effect of lamivudine dose-effect and aging relationship. Results The bands of absorbance (A) of HBV X gene in lamivudine group and control group were 151.4 ± 3.5 and 144.0 ± 11.4, respectively. The difference was not statistically significant. 4.36 × 104mol / L lamivudine sustained 24 hours can completely inhibit the X gene transcription, the control group and the treatment group (16 h) the purpose of the band A value were 243.9 ± 9.0 and 133.2 ± 7.8, P <0.01, The inhibitory effect with the drug concentration increases and the role of prolonged and enhanced. 4.36 × 104mol / L lamivudine continued role for 16 h, so that the maximum X-protein expression on behalf of the combined amount decreased from 353.3 ± 15.9 to 252.3 ± 18.8, P <0.01. Conclusion Lamivudine does not affect the replication of X gene in HepG2x, but significantly inhibits the transcription and protein expression of X gene.