体外分层渗透共培养研究胆管癌细胞株QBC939对正常内皮细胞的作用

Influence of human cholangiocarcinoma cells on normal endothelial cells

目的在体外分层渗透共培养体系中研究胆管癌细胞对正常内皮细胞的作用。方法建立胆管癌细胞株QBC939与正常内皮细胞的体外分层渗透共培养体系，以同期单独培养的正常内皮细胞为对照，用免疫荧光法检测内皮细胞与胆管癌细胞共培养后，内皮细胞Factin，pp125FAK，蛋白水解酶MMP-2、MMP-9、uPA蛋白表达变化，用Realtime PCR，Western印迹检测内皮细胞共培养前后，pp125FAK，MMP-2、MMP-9、uPA的mRNA及蛋白表达变化。结果与同期单独培养的正常内皮细胞对照相比，内皮细胞Factin表达增强，pp125FAK、MMP-2、MMP-9、uPA的mRNA及蛋白表达增加（P〈0．01）。结论胆管癌细胞能通过旁分泌作用导致内皮细胞骨架系统改变，并能诱导内皮细胞蛋白水解酶的基因表达及蛋白合成，表明肿瘤细胞对邻近内皮细胞还可以通过肿瘤细胞的内分泌、旁分泌等作用分泌各种细胞因子，作用于内皮细胞上的特异性受体，发挥其对内皮细胞mRNA表达的调控作用，而导致内皮细胞发生形态、行为的改变。

Objective To study the influence of human cholangiocarcinoma cells on normal endothelial cells in a transwells co-culture system. Methods Normal endothelium cells and human cholangioearcinoma cells of the line QBC939 transwells co-culture system were established. Normal endothelial cells cultured were used as control. Expression of F-actin, pp125FAK, MMP-2, MMP-9 and uPA was analyzed with immunofluorescent method. Expression of the genes pp125FAK, MMP-2, MMP-9 and uPA was analyzed by reabtime PCR and western blot. Results The expression of F-actin and the genes pp125FAK, MMP2, MMP-9 and uPA in the ECs co-cultured with QBC939 cholangiocarcinoma cells were all up-regulated in comparison with those in the controls （P〈0.01）. Conclusion Cholangiocarcinoma ceils promote the expression of proteolytic enzyme of ECs by paracrine action and induce the changes of ECs cytoskeleton system.