肝癌相关成纤维细胞对肝细胞肝癌影响的实验研究  

Effect of hepatocellular carcinoma associated fibroblasts to hepatocellular carcinoma

作　　者：华学锋[1] 李团结[2] 贾昌昌[1] 陈冠中[1] 郭宇[1] 邱东波 台艳 张琪 陈规划[1] 

机构地区：[1]中山大学附属第三医院岭南医院器官移植科,广州510530 [2]安徽医科大学第一附属医院普外科 [3]广东省肝脏疾病研究重点实验室

出　　处：《中华肝脏外科手术学电子杂志》2013年第3期50-53,共4页Chinese Journal of Hepatic Surgery(Electronic Edition)

基　　金：广东省自然科学基金(S2012040006483)

摘　　要：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 hepatocellular carcinoma （HCC） associated fibroblasts （hCAF） to the metastasis of HCC. Methods The hCAF and normal fibroblasts （NF） cells were cultured for 2 to 3 days and then stained by immunofluorescence. The expression of a-smooth muscle actin （α-SMA）, fibroblast surface protein （FSP）,fibroblast activation protein （FAP） and vimentin of hCAF and NF ceils were observed. The effect of hCAF to cell morphology change and invasion and metastasis of HCC were observed through co-culture and scratch test of hCAF and MHCC97L-green fluorescent protein（GFP）, HepG2-GFP HCC cells. Results The expression of c^-SMA, FSP, FAP and vimentin were all observed in the hCAF and NF cells. The expression level of ct-SMA and FAP in hCAF cells were obviously higher compared with the NF. The hCAF and HCC cells were co-cultured for 96 hours. The typical clustering growth of MHCC97L-GFP cells changed to disseminated growth. The polygon shape of epithelial cell of HepG2-GFP cell changed to spindle shape of interstitial cell. The scratch of MHCC97L-GFP, HepG2-GFP cells and hCAF cells after 5 days co-culture lessened obviously, compared with NF cells. Conclusion The hCAF cells can promote the transformation of epithelial cells into interstitial cells and enhance the invasion and metastasis of HCC cells.

关 键 词：成纤维细胞 癌 肝细胞 肿瘤转移 平滑肌肌球蛋白 膜蛋白质类 

分 类 号：R735.7[医药卫生—肿瘤]