论文部分内容阅读
目的采用体外重建基底膜细胞侵袭实验,研究口腔癌相关成纤维细胞(Carcinoma-associated fibroblasts,CAFs)侵袭力,用免疫组织化学法,观察CAFs对相关蛋白的表达。探讨CAFs在口腔鳞癌中的病理意义。方法采用组织块贴壁法培养颊部正常成纤维细胞(Normal Fibroblasts,NFs)和癌相关成纤维细胞CAFs,利用包被matrigel的transwell小室模拟天然基底膜的结构,利用培养基浓度梯度差诱导细胞穿膜并计数穿膜细胞数量,比较CAFs和NFs的侵袭力差异。采用免疫组化法,比较CAFs和NFs对基质金属蛋白酶-2(matrix metalloprotease-2,MMP-2)和成纤维细胞激活蛋白(fibroblast activation protein,FAP)表达差异。结果 CAFs穿膜的细胞数量显著高于NFS(P<0.05),差异具有统计学意义。CAFs对MMP-2和FAP的蛋白表达阳性,NFs表达为阴性。结论口腔CAFs的体外侵袭能力明显高于NFs,且能特异性地分泌MMP-2和FAP蛋白。这提示肿瘤间质的侵袭特性可随着上皮癌变而同步激活,CAFs参与肿瘤细胞外基质的重建,与肿瘤侵袭转移有关。
OBJECTIVE: To investigate the invasion of oral cancer-associated fibroblasts (CAFs) by in vitro reconstitution of basement membrane cells, and to observe the expression of related proteins in CAFs by immunohistochemistry. To investigate the pathological significance of CAFs in oral squamous cell carcinoma. Methods Normal fibroblasts (NFs) and CAFs of cancer-associated fibroblasts were cultured by tissue block adhesion method. The structure of natural basement membrane was simulated by transwell chambers coated with matrigel, and the cells were induced by the difference of medium concentration gradients The number of penetrating cells was counted and the differences in invasiveness of CAFs and NFs were compared. The expression of matrix metalloprotease-2 (MMP-2) and fibroblast activation protein (FAP) in CAFs and NFs were compared by immunohistochemistry. Results The number of CAFs penetrating the membrane was significantly higher than that of NFS (P <0.05). The difference was statistically significant. CAFs positive for MMP-2 and FAP protein expression, NFs expression was negative. Conclusion The oral CAFs in vitro invasive ability was significantly higher than NFs, and can specifically secrete MMP-2 and FAP protein. This suggests that the invasion of tumor stroma may be activated simultaneously with epithelial carcinogenesis. CAFs are involved in the reconstruction of extracellular matrix of tumor, which is related to tumor invasion and metastasis.