CAJ | 학술논문

目的研究THP-1来源的巨噬细胞对肝癌细胞HepG2的上皮间质化作用(epithelialmesenchymal transition,EMT),探讨肿瘤相关性巨噬细胞(tumor-associated macrophages,TAMs)在肝癌进展中的作用及机制.方法将THP-1来源的巨噬细胞与HepG2细胞共培养模拟肝癌相关微环境,观察共培养后HepG2细胞的形态变化.利用免疫荧光(immunofluorescence,IF)及Westernblot检测HepG2细胞与巨噬细胞共培养后E-cadherin表达的变化(E-cadherin表达缺失是EMT的标志).通过FlowCytomix检测分析THP-1细胞及THP-1来源的巨噬细胞培养上清中部分细胞因子表达量的差异.结果 HepG2细胞与THP-1来源的巨噬细胞共培养后,其细胞形态发生明显变化,由原来上皮细胞形态转化成为一种梭形的间质细胞形态.IF及Western-blot均显示HepG2细胞共培养后E-cadherin表达明显下调.THP-1细胞激活并分化为巨噬细胞后,白细胞介素IL-8及IL-1β表达量分别增加了40倍和20倍,P＜0.01；肿瘤坏死因子TNF-α表达量增加了8倍,P=0.056.结论巨噬细胞可诱导HepG2细胞发生EMT,该作用可能与其分泌的细胞因子IL-8及IL-1β和TNF-α增高有关.
목적 연구THP-1래원적거서세포대간암세포HepG2적상피간질화작용(epithelialmesenchymal transition,EMT),탐토종류상관성거서세포(tumor-associated macrophages,TAMs)재간암진전중적작용급궤제.방법 장THP-1래원적거서세포여HepG2세포공배양모의간암상관미배경,관찰공배양후HepG2세포적형태변화.이용면역형광(immunofluorescence,IF)급Westernblot검측HepG2세포여거서세포공배양후E-cadherin표체적변화(E-cadherin표체결실시EMT적표지).통과FlowCytomix검측분석THP-1세포급THP-1래원적거서세포배양상청중부분세포인자표체량적차이.결과 HepG2세포여THP-1래원적거서세포공배양후,기세포형태발생명현변화,유원래상피세포형태전화성위일충사형적간질세포형태.IF급Western-blot균현시HepG2세포공배양후E-cadherin표체명현하조.THP-1세포격활병분화위거서세포후,백세포개소IL-8급IL-1β표체량분별증가료40배화20배,P＜0.01；종류배사인자TNF-α표체량증가료8배,P=0.056.결론 거서세포가유도HepG2세포발생EMT,해작용가능여기분비적세포인자IL-8급IL-1β화TNF-α증고유관.
Objective This study investigates the epithelial-mesenchymal transition effects exerted on human liver cancer cells HepG2 by THP-1 derived macrophages.The roles of tumor-associated macrophages (TAMs) on liver cancer progression and its mechanisms were explored.Methods HepG2 cells were cultured with THP-1 derived macrophages to mimic the microenvironment of liver cancer.After the culture treatment,morphological changes of the liver cancer cells were observed.Decreased E-cadherin expression is a hallmark of epithelial-mesenchymal transition (EMT),and the Ecadherin protein variations in the HepG2 cells were detected by immunofluorescence (IF) and Westernblot.FlowCytomix was carried out to screen the cytokines in the supernanants of THP-1 cells and THP-1 derived macrophages.Results After culture with macrophages,HepG2 cells revealed a morphological change.These cells lacked epithelial morphology and became a spindle-like mesenchymal cell phenotype.Additionally,the E-cadherin protein expression was reduced dramatically as measured by IF and Western-blot.IL-8 and IL-1β expression in the supernatants were increased 40 and 20 times,respectively,after THP-1 cells were activated to macrophages (P＜0.01).TNF-α expression was increased 8 times (P =0.056).Conclusion THP-1 derived macrophages could induce EMT effects on HepG2 cells,which may relate to the increased secretion of IL-1β,IL-8 and TNF-α.