马兜铃酸Ⅰ致人上尿路上皮细胞MMP9、VEGF、E-cadherin、ERK的活性变化

Aristolochic Acid Ⅰ Induced Abnormalities in Expressions of MMP9,VEGF,E- cadherin and ERK 1 /2 Activation in Human Urothelium SV-HUC-1 Cells and in Upper Urothelial Ccarcinomas

目的探讨马兜铃酸Ⅰ(aristolochic acidⅠ,AAⅠ)诱导的可能癌变机制。分析AAⅠ能否诱导人尿路上皮细胞(SV-HUC-1)MMP9、VEGF、E-cadherin的表达。方法 SV-HUC-1细胞暴露于低浓度的AAⅠ(2.5、5.0、10μg/ml)48h,用蛋白免疫印迹(Western blot)法和荧光定量PCR分析MMP9、VEGF、E-cadherin的表达。此外用Western blot法比较马兜铃酸相关上尿路肿瘤(upper urothelial carcinomas associated with aristolochic acid nephropathy,AA-UUC)和非马兜铃酸相关上尿路肿瘤(nonAA-UUC)MMP9、VEGF、E-cadherin、PERK1/2蛋白表达变化。结果随着AAⅠ浓度的增加,MMP9、VEGF逐渐表达增加,而E-cadherin表达下降。然而,当SV-HUC-1细胞预先用ERK1/2信号通路抑制剂(U0126)作用1h,MMP9、VEGF蛋白的表达被抑制,而E-cadherin蛋白的表达恢复。和non-AA-UUC相比,AA-UUC的MMP9、VEGF、PERK1/2呈高表达,而E-cadherin呈低表达。结论低密度的AAⅠ可能通过激活SV-HUC-1细胞的ERK1/2信号通路使MMP9、VEGF、E-cadherin表达失调。马兜铃酸(AA)可能是上尿路肿瘤发生的原因之一。

Objective To investigate the mechanism of Aristolochic acid Ⅰ （AA Ⅰ）-induced carcinogenesis and whether AA Ⅰ induced the expressions of MMP9,VEGF,E-cadherin in human uroepithelial cells（SV-HUC-1）.Methods After SV-HUC-1 cells were treated with low concentration of AA Ⅰ （2.5,5.0,10.0μg/ml） for 48h,Western blotting and qRT-PCR were used to detect the expression of MMP9,VEGF and E-cadherin.What＇s more,we aslo used Western blotting to compare the expression of MMP9,VEGF,E-cadherin,PERK1/2 between upper urothelial carcinomas associated with Aristolochic acid Nephropathy（AA-UUC） and common upper urothelial carcinomas（non-AA-UUC）.Results When the concentration of AA Ⅰ increased,the expression of MMP9,VEGF increased but the E-cadherin decreased.However,when cells were pretreated with ERK1/2 signaling pathway inhibitors U0126 for 1h prior to exposure to AA Ⅰ,the effect of AA Ⅰ on expression of MMP9,VEGF levels was suppresssed but assisted the expression levels of E-cadherin.Morover,compared to non-AA-UUC,the expression of MMP9,VEGF,PERK1/2 on AA-UUC showd high but the expression of E-cadherin was low.Conclusion These findings suggest that low concentration of AA Ⅰ might induce MMP9,VEGF,E-cadherin dysregulation through ERK1/2 activation in SV-HUC-1 cells.AA might be associated with upper urinary tract urothelial carcinomas（UUC）.