肝癌细胞中AFP与RAR相互作用的实验研究
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摘要
目的探讨甲胎蛋白(alpha-fetoprotein,AFP)与维甲酸受体(retinoic acid receptor,RAR)之间的相互作用。方法采用免疫荧光、荧光能量共振转移(fluorescence resonance energy transfer,Fret)及免疫共沉淀实验验证肝癌细胞系Hep G2和SMMC-7721胞质中AFP与RAR之间的相互关系。结果在胞质中AFP与RAR存在共定位现象,Fret结果显示,两者之间存在很高的能量转移效率,分别为21%和19%。免疫共沉淀实验结果显示,胞质中AFP与RAR存在相互作用。结论 AFP与RAR存在相互作用,为后续研究AFP的生物学功能,以及维甲酸的药物疗效提供实验基础。
Objective To study the interaction of AFP and RAR. Methods In this study, hepatocellular carcinoma cells Hep G2 and SMMC-7721 were analyzed by confocal microscopy, fluorescence resonance energy transfer(Fret) and co-immunoprecipitation(Co IP) to verify the relationship of AFP and RAR. Results Confocal microscopy results showed that AFP co-localized with RAR in cytoplasm. And Fret showed that high energy transfer efficiency existed between AFP and RAR in Hep G2(21%) and SMMC-7721(19%), indicating these two proteins might be interacted with each other.Subsequent Co IP experimental results confirmed that in cytoplasm AFP could interact with RAR. Conclusion It is confirmed that the AFP could interact with RAR. And it has laid a solid foundation for the future study of AFP biological function, and for the retinoic acid drug efficacy study.
Objective To study the interaction of AFP and RAR. Methods In this study, hepatocellular carcinoma cells Hep G2 and SMMC-7721 were analyzed by confocal microscopy, fluorescence resonance energy transfer(Fret) and co-immunoprecipitation(Co IP) to verify the relationship of AFP and RAR. Results Confocal microscopy results showed that AFP co-localized with RAR in cytoplasm. And Fret showed that high energy transfer efficiency existed between AFP and RAR in Hep G2(21%) and SMMC-7721(19%), indicating these two proteins might be interacted with each other.Subsequent Co IP experimental results confirmed that in cytoplasm AFP could interact with RAR. Conclusion It is confirmed that the AFP could interact with RAR. And it has laid a solid foundation for the future study of AFP biological function, and for the retinoic acid drug efficacy study.
引文
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[14]王珊珊,林明华,欧阳雅博,等.表达谱基因芯片筛选肝癌细胞中ATRA下游基因的应用研究[J].河北医药,2016,38:2572-2574.
[2]Ko YS,Bae JH,Sinn DH,et al.The clinical significance of serum alpha-fetoprotein in diagnosing hepatocellular carcinoma in a health screening population[J].Korean J Gastroenterol,2017,69:232-238.
[3]Ma WJ,Wang HY,Teng LS.Correlation analysis of preoperative serum alpha-fetoprotein(AFP)level and prognosis of hepatocellular carcinoma(HCC)after hepatectomy[J].World J Surg Oncol,2013,11:212.
[4]王珊珊,李慧,李刚.甲胎蛋白的分子结构及生物学功能[J].世界华人消化杂志,2014,22:1487-1494.
[5]Li M,Li H,Li C,et al.Alpha fetoprotein is a novel protein-binding partner for caspase-3 and blocks the apoptotic signaling pathway in human hepatoma cells[J].Int J Cancer,2009,124:2845-2854.
[6]Zhu M,Guo J,Xia H,et al.Alpha-fetoprotein activates AKT/m TOR signaling to promote CXCR4 expression and migration of hepatoma cells[J].Oncoscience,2015,2:59-70.
[7]Li M,Li H,Li C,et al.Alpha-fetoprotein:a new member of intracellular signal molecules in regulation of the PI3K/AKT signaling in human hepatoma cell lines[J].Int J Cancer,2011,128:524-532.
[8]Dauphinee MJ,Mizejewski GJ.Human alpha-fetoprotein contains potential heterodimerization motifs capable of interaction with nuclear receptors and transcription/growth factors[J].Med Hypotheses,2002,58:453-461.
[9]Wang S,Jiang W,Chen X,et al.Alpha-fetoprotein acts as a novel signal molecule and mediates transcription of Fn14 in human hepatocellular carcinoma[J].J Hepatol,2012,57:322-329.
[10]Li P,Wang SS,Liu H,et al.Elevated serum alpha fetoprotein levels promote pathological progression of hepatocellular carcinoma[J].World J Gastroenterol,2011,17:4563-4571.
[11]Kanki K,Akechi Y,Ueda C,et al.Biological and clinical implications of retinoic acid-responsive genes in human hepatocellular carcinoma cells[J].J Hepatol,2013,59:1037-1044.
[12]Mcculloch D,Brown C,Iland H.Retinoic acid and arsenic trioxide in the treatment of acute promyelocytic leukemia:current perspectives[J].Onco Targets Ther,2017,10:1585-1601.
[13]Senoo H,Yoshikawa K,Morii M,et al.Hepatic stellate cell(vitamin A-storing cell)and its relative--past,present and future[J].Cell Biol Int,2010,34:1247-1272.
[14]王珊珊,林明华,欧阳雅博,等.表达谱基因芯片筛选肝癌细胞中ATRA下游基因的应用研究[J].河北医药,2016,38:2572-2574.