骨髓基质细胞介导的微环境对人肺腺癌A549细胞的作用

Effects of marrow stromal cell-mediated microenvironment on human lung adenocarcinoma A549 cells

目的:探讨肿瘤微环境中人骨髓基质细胞HS-5对人肺腺癌A549细胞的作用及机制。方法:采用HS-5细胞条件培养基(HS-5 cell-conditioned medium,HS-5-CM)处理A549细胞,分别通过MTT实验和划痕愈合实验观察A549细胞的活力和迁移能力,应用q PCR检测CX3C趋化因子受体1(CX3C chemokine receptor 1,CX3CR1)的表达;加入丝裂原活化蛋白激酶/细胞外信号调节激酶(mitogen-activated protein kinase/extracellular signal-regulated kinases,MAPK/ERK)通路抑制剂U0126后,应用Western blot法检测MAPK/ERK信号通路活化相关蛋白ERK及其磷酸化水平,划痕愈合实验检测抑制MAPK/ERK通路对HS-5-CM效果的影响,应用Western blot检测CX3CR1的表达情况。结果:HS-5-CM可以促进A549细胞的活力和迁移能力(P<0.01),同时A549细胞中CX3CR1的表达升高(P<0.05)。MAPK/ERK通路抑制剂U0126可以有效抑制HS-5-CM处理后MAPK/ERK通路的激活(P<0.01),抑制HS-5-CM促进A549细胞迁移的能力(P<0.01),同时下调CX3CR1的表达(P<0.05)。结论:HS-5细胞介导的微环境可以促进A549细胞的活力和迁移,其机制可能涉及MAPK/ERK信号通路的活化和CX3CR1的表达。

AIM:To investigate the effects of marrow stromal cell line HS-5 on human lung adenocarcinoma A549 cells in the tumor microenvironment.METHODS:The effects of HS-5 cell-conditioned medium(HS-5-CM)on the viability and migration ability of A549 cells were detected by MTT assay and wound-healing assay.After treatment with HS-5-CM,the expression of CX3C chemokine receptor 1(CX3CR1)at mRNA level in the A549 cells was examined by qPCR.The protein levels of p-ERK and ERK in the A549 cells treated with MAPK/ERK pathway inhibitor U0126 were observed by Western blot,the migration ability of the A549 cells was measured by wound-healing assay,and the protein expression of CX3CR1 was determined by Western blot.RESULTS:HS-5-CM promoted the viability and migration ability of the A549 cells(P<0.01).The expression of CX3CR1 at mRNA level in the A549 cells was increased after treatment with HS-5-CM.MAPK/ERK inhibitor U0126 inhibited the activation of MAPK/ERK signaling pathway(P<0.01),and reduced the migration ability(P<0.01)and the expression of CX3CR1(P<0.05)in the A549 cells.CONCLUSION:HS-5-CM significantly promotes the A549 cell viability and migration ability.Activation of MAPK/ERK signaling pathway and the expression of CX3CR1 may play a important role in this process.