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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 detect the expression of Prdx1 in lung cancer and to investigate the effect of Prdx1 on the formation of vasculoenic mimicry (VM) in lung cancer. Methods: The expression of VM and Prdx1 was detected by CD31 / PAS double staining and immunohistochemistry. The Prdx1 expression plasmid was transfected into lung cancer cell lines A549 and H1299 to induce exogenous up-down expression of Prdx1. Western blot was used to detect the expression of Prdx1, E-cadherin, VE-cadherin, P38MAPK, P38, P-P38 in A549-Prdx1 P38MAPK pathway inhibitor SB203580 was added into the cell line to detect the expression changes of P38, P-P38 and VEGF. The effects of Prdx1 on tube formation, migration and invasion were examined by three-dimensional, scratch and invasion assay. Results: Prdx1 was closely related to the formation of VM, lymph node metastasis and poor prognosis in patients. The expression of E-cadherin, Vimentin and VE-cadherin were up-regulated in Prdx1 and P38MAPK, respectively. The expression of P-P38 was up-regulated. Cell migration and invasion into the tube capacity was significantly enhanced. Contrary to the above results when Prdx1 is down-regulated. After P38MAPK pathway inhibitor SB203580 was added to A549-Prdx1 cell line, VEGF high P-P38 decreased with increasing dose, while P38 total protein remained unchanged. Conclusion: Prdx1 is highly expressed in lung cancer and may affect the formation of VM in patients with P38MAPK pathway.