探讨HMGB1对诱导人肺腺癌上皮细胞(A549)上皮间质转化的影响

To investigate the effect of HMGB1 on inducing epithelial mesenchymal transition in human lung adenocarcinoma epithelial cells (A549)

目的研究HMGB1对诱导人肺腺癌上皮细胞(A549)上皮间质转化的影响。方法通过不同浓度rHMGB1(0、0.5、1、2、4μg/mL)体外刺激A549细胞,western blot检测各浓度组vimentin蛋白相对表达量,得出rHMGB1体外刺激的最佳作用浓度。然后分三组实验,HMGB1组、TGF-β组(阳性对照)、control组(阴性对照),分别以2μg/mL rHMGB1、2μg/mL TGF-β、RPMI1640培养基体外刺激A549细胞。37℃、5%CO_(2)培养48 h或96 h后,MTT检测各组细胞增殖率、transwell检测各组细胞迁移能力,western blot检测各组细胞E-cadherin、vimentin和snail1蛋白相对表达量。结果HMGB1组和TGF-β组分别与control组比较,细胞增殖率显著增加(分别为P<0.0001和P<0.01);细胞迁移数量明显增加(P<0.0001);胞内snail1、vimentin蛋白相对表达量明显增加(P<0.0001),E-cadherin蛋白相对表达量明显降低(P<0.0001)。结论HMGB1体外有诱导人肺腺癌上皮细胞(A549)上皮间质转化的作用。

Objective To investigate the effect of HMGB1 on inducing epithelial mesenchymal transition in human lung adenocarcinoma epithelial cells(A549).Methods A549 cells were stimulated in vitro by different concentrations of rHMGB1(0,0.5,1,2,4μg/mL),and the relative expression of vimentin protein in each concentration group was detected by western blot assay,and the optimal in vitro action concentration of rHMGB1 was derived as 2μg/mL.Then the experiments were divided into three groups,HMGB1 group,TGF-β group(as positive control),control group(as negative control).2μg/mL rHMGB1,2μg/mL TGF-βand RPMI1640 medium only were used to stimulate A549 cells in vitro,separately.And after incubation at 37℃and 5% CO2 for 48 h or 96 h,MTT assay was performed to detect the proliferation rate of each group,transwell assay was performed to detect the migration ability of each group,and the relative expression of E-cadherin,vimentin and snail1 protein in each group was detected by western blot assay.Results The proliferation rate,the number of cell migration,the relative expression of intracellular snail1 and vimentin proteins were significantly increased(P<0.0001 or P<0.01)and the relative expression of E-cadherin protein was significantly decreased(P<0.0001)in A549 cells of the HMGB1group and the TGF-β group compared with the control group.Conclusion HMGB1 has the effect of inducing epithelial mesenchymal transformation of human lung adenocarcinoma epithelial cells A549 in vitro.