外源性HBx在昆明小鼠体内促进肝前体细胞上皮间质转化

发布时间：2018-11-20 06:57

【摘要】：目的构建靶向作用于小鼠肝前体细胞的动物模型,研究乙型肝炎病毒X蛋白(HBx)对肝前体细胞上皮间质转化(EMT)的影响。方法每周2次给予昆明小鼠2 m L/L四氯化碳灌胃,4周后行经门静脉注射稳定表达HBx的肝前体细胞和稳定表达空载体的肝前体细胞同时切除部分肝脏。术后继续每周2次进行灌胃,并分别于术后3、5、7、14、21、28 d,处死小鼠取肝脏标本;实时定量PCR检测小鼠肝组织HBx的mRNA水平、免疫组织化学染色检测肝组织中外源性细胞的存活。实时荧光定量PCR检测上皮钙黏素(E-cadherin)、神经钙黏素(N-cadherin)、波形蛋白(vimentin)和细胞角蛋白18(CK18)mRNA水平,Western blot法检测E-cadherin、N-cadherin、vimentin、CK18蛋白水平。结果免疫组织化学染色结果显示,术后肝脏组织明显有外源性细胞存活.随注射时间延长,肝组织HBx的含量增加,表达区域增大;实时荧光定量PCR和Western blot结果均显示过表达HBx能够使小鼠肝组织中E-cadherin、CK18水平降低,而N-cadherin、vimentin的水平增加。结论动物模型证实HBx在肝前体细胞EMT过程中起着重要的调控作用。
[Abstract]:Objective to study the effect of hepatitis B virus X protein (HBx) (HBV X protein (HBx) on the epithelial mesenchymal transformation (EMT) of liver precursor cells in mice. Methods Kunming mice were given 2 mL / L carbon tetrachloride intragastrically twice a week. After 4 weeks, liver precursor cells with stable expression of HBx and liver precursor cells with stable empty expression vector were injected through portal vein at the same time. The mice were sacrificed for 28 days after operation, the mRNA level of HBx in liver tissue was measured by real-time quantitative PCR, and the survival of exogenous cells in liver tissue was detected by immunohistochemical staining. E-cadherin (E-cadherin), neuronal cadherin (N-cadherin), vimentin (vimentin) (vimentin) and cytokeratin 18 (CK18) mRNA were detected by real-time fluorescence quantitative PCR. Results Immunohistochemical staining showed that exogenous cells survived obviously in liver tissue after operation. The results of real-time fluorescence quantitative PCR and Western blot showed that overexpression of HBx could decrease the level of E-cadherin CK18 and increase the level of N-cadherin vimentin in liver tissue of mice. Conclusion HBx plays an important regulatory role in the EMT process of liver precursor cells in animal models.
【作者单位】：重庆医科大学病理生理学教研室;重庆医科大学分子医学与肿瘤研究中心;
【基金】：国家自然科学基金青年项目(81201679)
【分类号】：R735.7

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