CAJ | 학술논문

目的：探讨肝癌细胞系HepG2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HepG2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HepG2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。
목적：탐토간암세포계HepG2화HLE중，AFP삼여RA－RAR신호통로적작용방식급기의의。방법Western blotting검측저량충세포중AFP여RAR적함량；ATRA처리세포후，면역형광법검측세포중RAR정위；류식세포술（ FCM）검측세포조망정황；채용격광공취초현미경관찰AFP여RAR재세포중적정위정황；면역공침정험증AFP여RAR적상호작용。결과 Western blotting결과현시HepG2세포위AFP양성이HLE위음성；면역형광결과현시가입ATRA후，RAR발생핵전위화핵취집현상；FCM결과현시ATRA능구인기저량충간암세포조망증가；격광공취초현미경결과현시AFP여RAR재포장중존재공정위현상；면역공침정결과증실재HepG2세포중AFP여RAR존재상호작용，이HLE인결실AFP이검측불도。결론 AFP통과억제RA－RAR신호통로적전체종이삼여조공간암적발생발전。
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 .