口腔癌相关成纤维细胞对人工基底膜的降解作用及其机制

Investigation of the Degradation and the mechanisms of Artificial Basement Membrane Co-cultured with Oral Carcinoma-associated Fibroblasts

目的观察口腔癌相关成纤维细胞(CAFs)降解人工基底膜matrigel的能力,并探讨其可能的作用机制。方法matrigel凝胶与CAFs和NFs共同培养24、48、72h后,收集培养上清液,按照羟脯氨酸检测试剂盒说明测定羟脯氨酸含量;用考马斯亮兰微孔板法行微量蛋白定量;用明胶酶谱分析对基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)含量进行测定。结果在各时间段,口腔CAFs上清超滤液中羟脯氨酸含量、上清液总蛋白浓度及MMP-2酶原含量明显高于NFs,且CAFs组表达大量MMP-2活性酶。口腔CAFs和NFs皆不表达MMP-9。结论CAFs具有较强的降解细胞外基质的能力,提示其与肿瘤-宿主微环境中基质重建及肿瘤侵袭有关。

Objective To investigate the degradation of artificial basement membrane （matrigel） co-cultured with oral carcinoma-associated fibroblasts （CAFs） and its possible mechanism. Methods CAFs and normal fibroblasts （NFs） were incubated on matrigel for 24, 48, 72 h. Equivalent amounts of conditioned medium were collected and assayed for total protein, hydroxyproline and matrix metalloproteinase-2（MMP-2）, matrix metalloproteinase-9 （MMP-9） activity by gelatin zymography. Results Oral CAFs were superior to oral NFs in total protein and hydroxyproline density, CAFs present more pro-MMP-2 and activated MMP-2. Conclusion CAFs were superior to NFs in degradation of matrigel. CAFs might play a key role in the reconstitution of extracellular matrix and the progression of tumor.