Effect of 1-phenyl 2-thiourea on the expressions of tumor multidrug resistance-related genes of Tg(abcb4:eGFP)zebrafish
Objective To investigate the effect of depigmentation agent 1-phenyl 2-thiourea(PTU)on the expressions of tumor multidrug resistance-related gene abcb4 and its labelling marker egfp of Tg(abcb4:eGFP)zebrafish and the possible underlying mechanism.Methods Twenty-four hpf(hour post fertilization)Tg(abcb4:eGFP)zebrafish embryos at 24 hours after fertilization were exposed to PTU,divided into control,PTU and PTU+RES groups.PTU treatment lasted for 96 hours.The fluorescent intensity of the gastrointestinal tract of zebrafish was examined in each group by taking the images under a fluorescent microscope.RT-qPCR was used to detect the mRNAs of abcb4,egfp,and pxr.Western Blot was applied to examine the protein expressions of abcb4 and egfp.Abcb4 function in different groups was assayed using Rhodamine B accumulation experiment.Results When compared to control group,the fluorescent intensity of Tg(abcb4:eGFP)zebrafish in PTU group was enhanced(P＜0.01),while the fluorescence change in PTU+RES group was not significant(P＞0.05).The mRNA expressions of abcb4,egfp,and pxr genes were increased in PTU group(P＜0.005).pxr mRNA expression was increased in PTU+RES group(P＜0.005).There were no significant changes in mRNA expressions of abcb4 and egfp genes(P＞0.05).The protein expressions of abcb4 and egfp were increased in PTU group(P＜0.005),while their expressions had no significant changes in PTU+RES group(P＞0.05).Abcb4 function was enhanced in PTU group(P＜0.01),while its function had no significant change in PTU+RES group(P＞0.05).Conclusion PTU can upregulate the expressions of abcb4 and egfp of Tg(abcb4:eGFP)zebrafish.Nuclear factor pxr may be involved in PTU-mediated induction and regulation of abc4 and egfp.When screening drugs in PTU-treated Tg(abcb4:eGFP)zebrafish,the attention should be paid to the effect of PTU on the expression of abcb4 and egfp.
1-phenyl 2-thioureatransgenic zebrafishzebrafish ATP-binding cassette transporter b4human ATP-binding cassette transporter b1pregnane X receptorenhanced green fluorescent pro-tein
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维普
