氯化钴致低氧诱导肺癌细胞系A549 DLL4表达上调并分泌促HUVEC迁移物

Up regulation of DLL4 expression in lung cancer A549 cells in hypoxia induced by CoCl_2 and secreted promote HUVEC migration material

目的探讨氯化钴(CoCl2)化学模拟低氧对肺癌细胞(A549)DLL4表达的影响及其与内皮细胞迁移的关系,为寻找抗肿瘤血管新生的潜在靶点提供理论依据。方法 A549分为对照组和CoCl2干预低氧组,EdU掺入法测定细胞增殖;用免疫荧光细胞染色和Western blot测定DLL4和低氧诱导因子HIF-1α在A549细胞中的表达;细胞划痕实验检测脐静脉内皮细胞(HUVEC)的迁移变化。结果低浓度氯化钴(200μmol/L)干预12 h后,A549细胞增殖没有明显抑制,DLL4和HIF-1α的表达均明显上调,低氧组DLL4蛋白表达为0.194±0.028显著高于对照组的0.098±0.015(P<0.05)。培养上清液促进HUVEC迁移。结论低氧明显上调A549细胞DLL4的表达,其对内皮细胞的迁移趋化作用提示其为抗肿瘤血管新生治疗的潜在靶点。

Objective To explore the expression of DLL4 in A549 after CoCl_2 treatment and elucidate the relationship between hypoxia induced factor-1α（HIF-1α） and delta like legend 4（DLL4）.Migration of human umbilical vein endothelial cells（HUVECs） in the condition of cancer culture supernatant was also observed.MethodsA549 cells were cultured and divided into 2 groups,control group and CoCl_2 treated group.Cell proliferation was detected by EdU incorporation assay.Expression of DLL4 and HIF-1α were detected by immunofluorescence staining and western-blot.Migration of endothelial cells was measured in a cell scratch wound model.Results The viability and proliferation of A549 were not significantly decreased after the CoCl_2（200 μmol/L） treatment.Expression of HIF-1α and DLL4 were increased in A549 after CoCl_2 treatment compared with normal condition.DLL4 expression was significantly higher in CoCl_2 treated group（0.194±0.028） compared with control（0.098±0.015）（P0.05）.Culture supernatant from CoCl_2 treated could enhance the migration of endothelial cells.Conclusion HIF-1α and DLL4 could be stimulated under hypoxic condition in A549.A549 in hypoxic condition can trigger the migration of endothelial cells,which is critical to angiogenesis.DLL4 might participate in tumor an-giogenesis and it is a promising target in tumor treatment.