【摘要】 目的探讨JWA基因在氯化高铁血红素(hemin)和热应激单独或联合作用下的表达规律。方法用Western blot方法检测JWA蛋白表达；用逆转录聚合酶链反应(RT-PCR)方法检测JWA mRNA表达变化；用氯霉素乙酰转移酶-酶联免疫吸附试验(CAT-ELISA)方法检测hemin和热应激对JWA基因近端启动子转录活性的影响。结果50μmol/L hemin处理K 562细胞1周可使JWA蛋白表达增高(3,23±0．57)倍；而30μmol/L hemin处理K 562细胞48 h可使JWA mRNA表达增加(1．37±0．06)倍。42℃处理K 562细胞2 h可使JWA蛋白表达增高(8．00±1．73)倍；42℃处理K 562细胞30 min可使JWA mRNA表达明显升高(1．87±0．13)倍。hemin和热应激联合作用于K 562细胞后,JWA蛋白和mRNA的表达均低于单独处理。CAT-ELISA结果显示hemin及热应激单独处理可上调各自反应元件的应答,但二者联合作用后反应元件的应答似乎呈现一种部分拮抗的作用。结论hemin和热应激可能分别通过JWA近端启动子上各自的反应元件而调节JWA基因的表达。更多还原

【Abstract】 Objective To investigate the expression of JWA after heroin and(or)thermal stress exposure. Methods The expression of JWA was determined by Western blot.RT-PCR was carried out for evaluation of the expression of JWA mRNA.The JWA promoter transcription activity analysis was performed by CAT-ELISA. Results The expression of JWA protein was significantly increased by 3.23±0.57 times of control in K562 cells after treated by heroin(50μmol/L)for 1 week,and the similar pattern was observed in the cells after treatment with thermal stress(42℃)for 2 hours(increased by 8.00±1.73 times).The expression of JWA mRNA was also signif- icantly elevated by 1.37±0.06 times in K562 cells treated by heroin(30μmol/L)for 48 hours,and with a similar regulation pattern(increased by 1.87±0.13 times)by treatment with thermal stress(42℃)for 30 minutes.How- ever,an antagonistic effect was observed by treatment of K562 cells with hemin(30μmol/L,48 hours)followed by thermal stress(42℃,30 minutes).The CAT-ELISA further confirmed that hemin or thermal stress treatment alone up-regulated JWA transcription activity while the effects could be counteracted partly by the combined treatment of the both.Conclusion The hemin and thermal stress may regulate JWA expression via distinct intracellular signal transduction pathways.更多还原