Prdx1通过P38MAPK通路影响肺癌VM的形成

Prdx1 promotes vasculogenic mimicry formation of lung cancer via P38MAPK signaling

目的:检测Prdx1在肺癌中的表达,探讨Prdx1对肺癌血管生成拟态(vasculoenic mimicry,VM)形成的影响,并深入研究其影响机制。方法:利用CD31/PAS双重染色及免疫组织化学法检测并分析VM及Prdx1的表达关系。将Prdx1表达质粒转染至肺癌细胞系A549、H1299中,诱导Prdx1外源性升降表达。Western blot检测转染前、后Prdx1、EMT相关蛋白(E-cadherin、Vimentin)、VM相关蛋白(VE-cadherin、VEGF)、P38MAPK通路相关蛋白(P38、P-P38)表达变化情况;A549-Prdx1细胞系加入P38MAPK通路的抑制剂SB203580后,检测(P38,P-P38、VEGF)的表达变化情况;三维,划痕和侵袭实验检测Prdx1对细胞成管、迁移、侵袭能力的影响。结果:Prdx1与患者VM的形成、淋巴结转移、不良预后密切相关。Prdx1升表达高E-cadherin表达下调,Vimentin表达上调,VE-cadherin、VEGF表达上调,P38MAPK中P38表达不变,P-P38表达上调。细胞的迁移侵袭成管能力显著增强。Prdx1低表达后与上述结果相反。A549-Prdx1细胞系中加入P38MAPK通路抑制剂SB203580后,VEGF高P-P38随剂量的增加而降低,而P38总蛋白不变。结论:Prdx1在肺癌中高表达,可能通过P38MAPK通路影响患者VM的形成。

Objective： To analyze the effect of Prdxl on lung cancer by investigating its expression and role in vasculogenic mimicry （VM） formation. Methods： The relationship between VM existence and Prdxl expression was detected and analyzed by CD31/PAS du- al staining and immunohistochemical staining. A549 and H1299 cells were transfected with Prdxl-expressing plasmids to induce exoge- nous Prdxl protein expression. Changes in the expression levels of Prdxl, EMT-related proteins （E-cadherin and Vimentin）, VM-related proteins （VE-cadherin and VEGF）, and P38MAPK signaling-related proteins （P38 and P-P38） were detected by Western blot after trans- fection. Furthermore, changes in the expression levels of P38MAPK signaling-related proteins （P38 and P-P38） and VM-related protein （VEGF） in A549-Prdxl cells were detected by Western blot after using SB203580. The effects of Prdxl gene transfection on migration capacity were determined by an in vitro wound-healing assay, whereas the role of Prdxl transfection in invasive potential was deter- mined by an invasion assay. The role of Prdxl transfection on tube structure formation potential was determined by three-dimensional culture. Results： Immunohistochemistry staining showed that Prdxl expression was positively associated with VM, metastasis, and poor prognosis. Western blot showed that after Prdxl was increased, E-cadherin expression was downregulatecl, whereas Vimentin, VE-cadherin, VEGF, and P-P38 expression levels were upregulatecl in A549 cells. Moreover, P38 expression was unchanged. Wound healing, invasion, and three-dimensional culture assays showed that the migration, invasion, and tube structure formation potentials of A549 cells significantly increased, whereas suppressing Prdxl in H1299 cells yielded the opposite results. After adding the P38MAPK pathway inhibitor, VEGF and P-P38 expression levels decreased as inhibitor dosage increased, whereas P38 expression remained the same. Conclusion： Prdxl, which is highly expressed in lung cancer specimens, was closely associated with ViVI. This association may promote VM formation via P38MAPK signaling.