胶原三维立体培养模型中胚肺成纤维细胞与肺癌GLC-82细胞的相互作用

Interaction between lung fibroblast and lung cancer GLC-82 cells in a three-dimensional collagen culture model

背景:组织细胞、炎细胞可能在肿瘤细胞的侵袭、转移中发挥着重要作用,而细胞间的相互作用对基质金属蛋白酶有影响。目的:观察胚肺成纤维细胞与肺癌GLC-82细胞在胶原三维立体培养条件下,相互作用对基质金属蛋白酶1,2,9表达的影响。方法:采用三维胶原立体培养模型,按比例混合胶原,4倍DMEM和灭菌水,1倍DMEM的液体胶原,其中胶原终浓度为0.75g/mL。将GLC-82细胞、胚肺成纤维细胞分别单独培养及GLC-82细胞和胚肺成纤维细胞按5∶1混合培养,待胶原固化后加入1倍DMEM培养基,48h后收集上清液。Western blot法检测基质金属蛋白酶1表达水平,明胶酶谱法测定基质金属蛋白酶2,9的表达。结果与结论:胚肺成纤维细胞与肺癌GLC-82细胞混合培养组基质金属蛋白酶1及基质金属蛋白酶2,9分泌量大于单独培养组(P<0.05)。结果提示,三维立体培养条件下,胚肺成纤维细胞与肺癌GLC-82细胞相互作用能通过上调基质金属蛋白酶1,2,9的表达和活化,促进肺癌侵袭和转移。

BACKGROUND：The histocyte and inflammatory cells played an important role in the invasion and metastasis of lung cancer cells.However,cell interaction influenced matrix metalloproteinase.OBJECTIVE：To observe the influence of interaction between fetal lung fibroblasts（HFL） and lung cancer cells on matrix metalloproteinase-1,2,and 9（MMP-1,2,9） expression in three-dimensional co-culture.METHODS：Human lung cancer cells（GLC-82） and HFL were co-cultivated in three-dimensional collagen gels,which were mixed with collagen,4 time volume DMEM and sterilizing water,as well as 1 time volume DMEM.The final concentration of collagen was 0.75 g/mL.GLC-82 and HFL were cultured individually,and GLC-82 and HFL were then co-cultured according to the ratio of 5：1.After 48 hours,the supernatant was harvested following adding 1 time volume DMEM.MMP-1 expression was detected by Western blot and MMP-2 and 9 expressions were detected by gelatin zymography.RESULTS AND CONCLUSION：In three-dimensional co-culture of GLC-82 with HFL,the expressions of MMP-1,MMP-2 and MMP-9 were much higher than that in GLC-82 or in HFL alone（P〈0.05）.This suggested that the interaction between HFL and lung cancer cells enhanced the invasion and metastasis of lung cancer through up-regulating the secretion and activation of MMP-1,MMP-2,and MMP-9.