1-苯基2-硫脲对Tg(abcb4:eGFP)斑马鱼肿瘤多重耐药相关基因表达的影响  

Effect of 1-phenyl 2-thiourea on the expressions of tumor multidrug resistance-related genes of Tg(abcb4:eGFP)zebrafish

作　　者：孔鹏 莫大双 陈敏 黄江涛 安丽娟 尹正昊 舒莉萍 何志旭 KONG Peng;MO Dashuang;CHEN Min;HUANG Jiangtao;AN Lijuan;YIN Zhenghao;SHU Liping;HE Zhixu(Department of Pediatrics,School of Clinical Medicine,Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Children's Emergency,Guizhou Branch of Shanghai Children's Medical Center Shanghai Jiaotong University School of Medicine,Guiyang 550004,Guizhou,China;Department of Immunology,School of Basic Medicine,Guizhou Medical University,Guiyang 550004,Guizhou,China;Key Laboratory of Adult Stem Cell Translational Research,Chinese Academy of Medical Science,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学临床医学院儿科学教研室,贵州贵阳550004 [2]上海交通大学医学院附属上海儿童医学中心贵州医院儿童急诊部,贵州贵阳550004 [3]贵州医科大学基础医学院免疫学教研室,贵州贵阳550004 [4]中国医学科学院成体干细胞转化研究重点实验室,贵州贵阳550004

出　　处：《贵州医科大学学报》2024年第11期1587-1592,1600,共7页Journal of Guizhou Medical University

基　　金：国家自然科学基金(32160168,322260177);贵州省科技计划项目(黔科合平台人才CXTD[2021]002)。

摘　　要：目的探讨去色素药物1-苯基2-硫脲(PTU)对Tg(abcb4:eGFP)斑马鱼肿瘤多重耐药相关基因abcb4及其标记基因egfp表达的影响及可能机制。方法选取受精后24 h(24 hpf)Tg(abcb4:eGFP)斑马鱼胚胎做药物暴露实验,分为Control、PTU以及PTU+RES 3个组;持续用药96 h,通过荧光显微镜拍照检测各组斑马鱼胃肠道荧光强度;RT-qPCR检测abcb4、egfp及pxr基因mRNA,Western Blot检测abcb4、egfp蛋白(Abcb4、eGFP)的表达,罗丹明B积累实验检测不同组Abcb4功能。结果与Control组相比,PTU组Tg(abcb4:eGFP)斑马鱼荧光表达增强(P<0.01),PTU+RES组荧光变化不明显(P>0.05);PTU组abcb4、egfp及pxr基因mRNA表达均增加(P<0.005),PTU+RES组pxr基因mRNA表达增加(P<0.005),abcb4、egfp基因mRNA表达无明显变化(P>0.05);PTU组abcb4、egfp蛋白表达均增加(P<0.005),PTU+RES组无明显变化(P>0.05);PTU组Abcb4功能增强(P<0.01),PTU+RES组无明显变化(P>0.05)。结论PTU可诱导Tg(abcb4:eGFP)斑马鱼abcb4和egfp表达增加,核因子pxr可能参与PTU对abcb4和egfp的诱导调节过程,在应用PTU去色素处理的Tg(abcb4:eGFP)斑马鱼进行药物筛选时需注意PTU对abcb4和egfp表达的影响。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.

关 键 词：1-苯基2-硫脲 转基因斑马鱼 斑马鱼b4型ATP结合盒 人B1型ATP结合盒 孕烷X受体 增强型绿色荧光蛋白 

分 类 号：R73-37[医药卫生—肿瘤]