癌相关成纤维细胞对结肠癌生物学行为的影响  

Carcinoma - associated fibroblasts effect on biological behavior of colon carcinoma LoVo cells in vivo

作　　者：施琳[1] 隋贵春 袁宏伟[1] 韦玄[5] 贾永峰[1] 云芬 孙红[4] 乌新林[2] 

机构地区：[1]内蒙古医科大学附属医院病理科,呼和浩特010059 [2]内蒙古医科大学附属医院普通外科,呼和浩特010059 [3]内蒙古医科大学,呼和浩特010110 [4]内蒙古医科大学基础医学院生物医学综合实验中心,呼和浩特010110 [5]中国药科大学基础医学与临床药学学院,南京210098

出　　处：《中华实验外科杂志》2016年第7期1780-1784,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（81260364）;教育部春晖计划（Z2012007）;内蒙古自然科学基金（2012MS1123、2013MS1132）;内蒙古白治区高等学校青年科技英才支持计划（NJYT-15-A10）

摘　　要：目的观察人结肠癌相关成纤维细胞（CAFs）在体内对结肠癌LoVo细胞侵袭转移的影响。方法胶原酶消化法原代培养人结肠CAFs和正常成纤维细胞（NFs）；对第3代人结肠CAFs和NFs通过形态学观察和免疫细胞化学染色方法进行鉴定；BALB／c裸鼠随机分3组，实验组将CAFs和NFs分别与结肠癌LOVo细胞按浓度为1×10^10／L，以1：1比例混合，以单独注射LoVo细胞为对照组，通过皮下注射和脾脏注射的方法分别建立皮下移植瘤模型和肝转移模型，比较3组间皮下移植瘤的牛长速度及肝转移率之间的差异；采用免疫组织化学链霉菌抗生物素蛋白-过氧化物酶连结（SP）法检测3组皮下移植瘤标本中CD31标记的微血管密度（MVD）以探讨CAFs和NFs对肿瘤血管生成的影响。结果CAFs较NFs特异表达成纤维细胞活化蛋白（FAP）和成纤维细胞特异性蛋白（FSP），两种细胞结蛋白（Desmin）均为阴性表达；CAFs＋LoVo组（CAF干扰组）肿瘤生长速度和体积明显大于NFs＋LoVo组（NF干扰组）和LoVo组（未干扰组，P〈0．05）；皮下移植瘤CD31免疫组织化学染色显示CAFs＋LoVo组MVD计数为（11．16±2．44）个，NFs＋LoVo组为（7．36±1．52）个，LoVo组为（6．08±1．41）个，CAFs＋LoVo组MVD计数明显多于NFs＋LoVo组和LoVo组（P〈0．05），NFs＋LoVo组MVD计数多于LoVo组（P〈0．05）；CAFs＋LoVo组肝转移率为80％，明显高于NFs＋LoVo组（40％）和LoVo组（20％，P〈0．05）。结论人结肠CAFs能够促进结肠癌的侵袭和转移，可能与CAFs促进结肠癌肿瘤血管生成有关。Objective To investigate the effects of human colon carcinoma - associated fibroblasts （CAFs） on colon cancer cells LoVo invasion and metastasis in vivo. Methods The primary human colon CAFs and normal fibroblasts （NFs） were obtained by digestion methods. Identified the third generation CAFs and NFs cells by morphological observation and immunocytochemistry （ICC） staining. BALB/c nude mice were randomly grouped. Mix CAFs or NFs with colon cancer LoVo cells at a concentration of 1×10^10 cells/L in a 1：1 ratio and inject alone LoVo cells as the control group, and these cells are injected into spleen and subcutaneous to establish the subcutaneous xenotransplanted tumor model and liver metastasis model. Compare the differences of the growth rate in subcutaneous tumors and liver metastasis rate in liver metastasis model among the three groups. Detection of microvessel density （MVD） labeled CD31 by immu- nohistochemical streptavid - in - peroxidase （SP） method in the xenograft specimens to investigate the role of CAFs in tumor angiogenesis. Results ICC shows that FAP, FSP, Desmin are negative in NFs; Desmin is negative, fibrohlast activiation protein （FAP） and fibroblast specific protein （FSP） are positive in CAFs. The tumor volume of CAFs ＋ LoVo group is significantly greater than those of NFs ＋ LoVo or LoVo group （ P 〈 0. 05 ）. Immunohistochemistry shows the MVD count of CAFs ＋ LoVo group are 11.16 ± 2. 44, NFs＋ LoVo group are 7. 36 ± 1.52, LoVo group are 6. 08 ± 1.41, MVD count of CAFs ＋ LoVo group are significantly more than of the NFs ＋ LoVo and LoVo group （ P 〈 0. 05 ）. The liver metastasis rate of CAFs ＋ LoVo group （80%） is, significantly higher than NFs ＋ LoVo （40%） or LoVo group （20%, P 〈0. 05）. Conclusion CAFs can promote human colon tumor angiogenesis. CAFs might promote human colon cancer invasion and metastasis.

关 键 词：结肠癌 癌相关成纤维细胞 肝转移 皮下移植瘤 

分 类 号：R737.25[医药卫生—肿瘤]