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中华肝脏外科手术学电子杂志 ›› 2013, Vol. 02 ›› Issue (03) : 194 -199. doi: 10.3877/cma.j.issn.2095-3232.2013.03.012

所属专题: 文献

基础研究

肝癌相关成纤维细胞对肝细胞肝癌影响的实验研究
华学锋1, 李团结2, 贾昌昌1, 陈冠中1, 郭宇1, 邱东波3, 台艳3, 张琪3, 陈规划1,()   
  1. 1. 510530 广州,中山大学附属第三医院岭南医院器官移植科
    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
  • 收稿日期:2013-04-01 出版日期:2013-06-10
  • 通信作者: 陈规划
  • 基金资助:
    广东省自然科学基金(S2012040006483)

Effect of hepatocellular carcinoma associated fibroblasts to hepatocellular carcinoma

Xue-feng HUA1, Tuan-jie LI2, Chang-chang JIA1, Guan-zhong CHEN1, Yu GUO1, Dong-bo QIU3, Yan TAI3, Qi ZHANG3, Gui-hua CHEN1,()   

  1. 1. Organ Transplantation Center, Lingnan Hospital, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510530, China
  • Received:2013-04-01 Published:2013-06-10
  • Corresponding author: Gui-hua CHEN
  • About author:
    Corresponding author: CHEN Gui-hua, Email:
引用本文:

华学锋, 李团结, 贾昌昌, 陈冠中, 郭宇, 邱东波, 台艳, 张琪, 陈规划. 肝癌相关成纤维细胞对肝细胞肝癌影响的实验研究[J]. 中华肝脏外科手术学电子杂志, 2013, 02(03): 194-199.

Xue-feng HUA, Tuan-jie LI, Chang-chang JIA, Guan-zhong CHEN, Yu GUO, Dong-bo QIU, Yan TAI, Qi ZHANG, Gui-hua CHEN. Effect of hepatocellular carcinoma associated fibroblasts to hepatocellular carcinoma[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2013, 02(03): 194-199.

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 α-smooth muscle actin(α-SMA), fibroblast surface protein (FSP), fibroblast activation protein (FAP) and vimentin of hCAF and NF cells 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 α-SMA, FSP, FAP and vimentin were all observed in the hCAF and NF cells. The expression level of α-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.

图1 荧光显微镜观察hCAF和NF的α-SMA、FSP、FAP、波形蛋白表达图像
图2 荧光显微镜观察hCAF、NF和MHCC97L-GFP共培养96 h图像
图3 荧光显微镜观察hCAF、NF和HepG2-GFP共培养96 h图像
图4 荧光显微镜观察hCAF与MHCC97L-GFP共培养5 d后划痕实验图像
图5 荧光显微镜观察HepG2-GFP与hCAF共培养5 d后划痕实验图像
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