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目的探讨肝癌相关成纤维细胞(hCAF)对肝细胞肝癌(肝癌)转移的影响。方法分别将hCAF和正常成纤维细胞(NF)培养2~3 d,行免疫荧光染色,观察hCAF和NF细胞中α-平滑肌肌动蛋白(α-SMA)、成纤维细胞表面蛋白(FSP)、成纤维细胞活化蛋白(FAP)、波形蛋白(vimentin)的表达情况。通过hCAF与MHCC97L-绿色荧光蛋白(GFP)、HepG2-GFP肝癌细胞共培养及划痕实验,观察hCAF对肝癌细胞的形态变化的影响及其对肝癌侵袭转移能力的作用。结果 hCAF和NF细胞均表达α-SMA、FSP、FAP、vimentin,但hCAF表达α-SMA与FAP的水平明显高于NF。hCAF与肝癌细胞共培养至96 h,MHCC97L-GFP细胞生长由典型的抱团样成簇生长,变成分散呈播散样生长;HepG2-GFP细胞由多角形的上皮细胞形态转变为长梭形的间质细胞形态。MHCC97L-GFP、HepG2-GFP肝癌细胞与hCAF共培养5 d后划痕明显较NF组缩小。结论 hCAF能加速肝癌细胞发生类似上皮细胞间质转型的变化,增强肝癌细胞侵袭转移能力。
Objective To investigate the effect of hCAF on the metastasis of hepatocellular carcinoma (HCC). Methods The hCAF and normal fibroblasts (NFs) were cultured for 2 ~ 3 days respectively. The expression of α-SMA, FSP, Fibroblast activation protein (FAP), vimentin (vimentin) expression. The hCAF and MHCC97L-GFP and HepG2-GFP hepatocarcinoma cells were co-cultured and scratched to observe the effect of hCAF on the morphological changes of hepatoma cells and its effect on the invasion and metastasis of HCC. Results Both α-SMA, FSP, FAP and vimentin were expressed in hCAF and NF cells, but the expression of α-SMA and FAP in hCAF was significantly higher than that in NF. hCAF cells were co-cultured with hepatocarcinoma cells for 96 h. The growth of MHCC97L-GFP cells was typical cluster-like clusters, and dispersed into disseminated samples. HepG2-GFP cells changed from polygonal epithelial cells to long fusiform Interstitial cell morphology. Scratches of MHCC97L-GFP and HepG2-GFP hepatocellular carcinoma cells co-cultured with hCAF for 5 days were significantly smaller than those of NF group. Conclusions hCAF can accelerate the change of epithelial-mesenchymal transition in hepatoma cells and enhance the invasion and metastasis of hepatocellular carcinoma cells.