阿霉素对模式生物斑马鱼中abcb4基因表达的影响

孙琮杰; 何志旭; 舒莉萍等

阿霉素对模式生物斑马鱼中abcb4基因表达的影响

Effect of Doxorubicin on abcb4Gene Expression in Zebrafish (Danio rerio) Embryos

摘要

摘要: 目的选择斑马鱼作为实验对象，研究阿霉素(doxorubicin，DOX)对其abcb4基因表达的影响，进一步了解abcb4基因在斑马鱼多药耐药机制中可能的作用。方法分别以2 mL/L二甲基亚砜（DMSO），10 μmol/L DOX及含2 mL/L DMSO的10 μmol/L DOX对斑马鱼胚胎进行药物处理，另以Eggwater处理的胚胎作为对照组。将正常发育的4~16个细胞期斑马鱼胚胎，随机分入以上各组中，药物处理至120 hpf。收集药物处理下的斑马鱼不同时期胚胎运用实时荧光定量PCR和胚胎原位杂交技术，检测abcb4基因在斑马鱼中的表达变化情况。结果相较于对照组，药物处理的斑马鱼胚胎abcb4基因mRNA表达水平升高(Pabcb5基因mRNA表达情况则无明显变化。通过斑马鱼胚胎原位杂交，均在斑马鱼120 hpf胚胎小肠部位发现有abcb4基因阳性杂交信号，且药物处理的斑马鱼胚胎在脑及心脏部位发现abcb4基因阳性杂交信号。结论 DOX能诱导斑马鱼胚胎abcb4基因表达水平增高，对阐明abcb4基因在斑马鱼多药耐药产生机制中的作用具有重要意义。

Abstract: Objective To determine the effect of doxorubicin (DOX) on abcb4gene expression and the role of abcb4gene in multidrug-resistance. Methods Zebrafish embryos were treated with 2 mL/L DMSO, 10 μmol/L DOX and 2 mL/L DMSO+10 μmol/L DOX, respectively. The zebrafish embryos treated with Eggwater served as controls. Exposures started at 4 to 16 cell stage of the embryos and terminated 120 hours post fertilization (hpf). The expression of abcb4gene in zebrafish embryos was examined on 48, 72, 96, and 120 hpf with whole-mount in situ hybridization (WISH) and quantitative real-time PCR (qPCR). Results Compared with the controls, DOX-exposed embryos had higher level of abcb4gene expression (Pabcb5gene. WISH showed that abcb4gene was expressed in the guts of zebrafish embryos. However, those exposed to DOX also showed strong WISH signals in the brain and heart. Conclusion Doxorubicin increases the expression of abcb4gene in zebrafish embryos. abcb4gene may play an important role in multidrug-resistance.

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