AFP抑制肝癌细胞中RA-RAR信号通路的传递  

Inhibitory effect of AFP on RA-RAR signal pathway in liver cancer cells

作　　者：王珊珊[1] 陈宁[1] 庞丽君[1] 欧阳雅博[1] 李刚[2] 陈德喜[1] 

机构地区：[1]首都医科大学附属北京佑安医院,北京市肝病研究所,北京市100069 [2]北京大学医学部基础医学院生物化学与分子生物学系

出　　处：《河北医药》2014年第24期3693-3695,共3页Hebei Medical Journal

基　　金：国家自然科学基金资助项目(编号:81370522);国家科技支撑计划项目(编号:2012BAI15B08);北京市肝病研究所所内基金项目(编号:BJIH-01402)

摘　　要：目的探讨肝癌细胞系Hep G2和HLE中,AFP参与RA-RAR信号通路的作用方式及其意义。方法Western blotting检测这两种细胞中AFP与RAR的含量;ATRA处理细胞后,免疫荧光法检测细胞中RAR定位;流式细胞术(FCM)检测细胞凋亡情况;采用激光共聚焦显微镜观察AFP与RAR在细胞中的定位情况;免疫共沉淀验证AFP与RAR的相互作用。结果 Western blotting结果显示Hep G2细胞为AFP阳性而HLE为阴性;免疫荧光结果显示加入ATRA后,RAR发生核转位和核聚集现象;FCM结果显示ATRA能够引起这两种肝癌细胞凋亡增加;激光共聚焦显微镜结果显示AFP与RAR在胞浆中存在共定位现象;免疫共沉淀结果证实在Hep G2细胞中AFP与RAR存在相互作用,而HLE因缺失AFP而检测不到。结论 AFP通过抑制RA-RAR信号通路的传递从而参与调控肝癌的发生发展。Objective To investigate the effect ,type of action and significance of AFP on RA-RAR signal pathway in HepG2 and HLE cells.Methods The expression levels of AFP and RAR in HepG 2 and HLE cells were detected by Western Blotting;after treated by ATRA for 6 hours,RAR location in HepG2 and HLE cells was detected by immunofluorescence;the cell apoptosis was tested by flow cytometry （ FCM ）;the location AFP and RAR in cells was observed by laser confocal microscopy;the interaction between AFP and RAR was validated by co-immunoprecipitation test .Results Western Blotting showed that AFP expression was positive in HepG 2 cells, however , which was negative in HLE cells .Immunofluorescence results showed that nuclear translocation and nuclear aggregation of RAR were found after ATRA treatment ;FCM analysis showed that ATRA could induce cells apoptosis of both HepG 2 and HLE cells;laser confocal microscopy showed that co-localized phenomenon was observed in AFP and RAR;co-immunoprecipitation results showed that there was a interaction between AFP and RAR in HepG2 cells,however,which was not found in HLE cells because of AFP deletion .Conclusion AFP is involved in the pathogenesis and development of hepatocellular carcinoma through inhibiting RA -RAR signal transduction pathway .

关 键 词：肝癌 AFP RAR 信号通路 

分 类 号：R735.7[医药卫生—肿瘤]