| Purpose:Vasculogenic mimicry(VM)is a special structure which surrounded by tumor cells,it does not depend on the endothelial cells and it is a special microcirculation model.VM is closely related with tumor invasion,metastasis,the survival time of patients and poor prognosis.Prdx1 is a member of the PrdxS family and Prdx1 plays an important role in tumor cell invasion and metastasis.In this study,we investigated the role of Prdx1 in the formation of VM in lung cancer and expected to provide a new therapeutic basis treatment strategy for the treatment of lung cancer.Methods:1?The expression of Prdx1,P38 and VEGF in lung cancer was observed by immunohistochemistry.The structure of VM in lung cancer tissues was observed by CD31 / PAS double staining.The correlation between Prdx1 and P38,VEGF and VM was analyzed statistically.Prdx1 was divided into Prdx1 positive group and Prdx1 negative group.The relationship between Prdx1 with sex,age,tumor size,TNM stage and lymph node metastasis was analyzed.Kaplan-Meier survival analysis was used to compare the survival prognosis of Prdx1 and VM positive patients.2?According to the expression level of Prdx1,the lung cancer cell line A549 with a low expressing and the lung cancer cell line H1299 with a high expression of Prdx1 were screened out.Then the A549 cell line was overexpressed and the H1299 cell line was downexpressed.And the stable transfected cell lines were screened out.The transfection effect of Prdx1 was assessed by Western Blot.3?The effects of migration and invasion of A549 cells were observed by scratch and invasion experiments.And the effect of H1299 cell migration and invasion was also observed after Prdx1 expression.4?The changes of tubular structure-forming ability of A549 cells after Prdx1 expression were analyzed by three-dimensional culture experiment.On the contrary,Prdx1 was observed to value the changes of tubular structure formation ability of H1299 cells.5?The effect of Prdx1 on the expression of EMT-related protein(E-cadherin and Vimentin)in A549 cells was detected by Western Blot.The effect of Prdx1 on the expression of EMT-related protein(E-cadherin and Vimentin)in H1299 cells was also detected by Western Blot.6?Western blot were used to detect the VM-related protein(VE-cadherin and VEGF)in A549 cells after Prdx1 upregulation and in H1299 cells after Prdx1 downregulation.7?Western blot were used to detect the P38MAPK-related protein P38 and P-P38 in A549 cells after Prdx1 upregulation and in H1299 cells after Prdx1 downregulation.8?After used SB203580,Western blot were used to detected the change of P38,P-P38 and VEGF in A549 cells after Prdx1 upregulation.Results:1?The results of immunohistochemical staining and CD31 / PAS double staining showed that the positive expression of Prdx1 was 64.36%(65/101),the positive expression of P38 was 71.29%,the positive expression of VEGF was 65.35%(66/101)in 101 cases of lung cancer and the positive expression of VM was 23.76%(24/101).Prdx1 was divided into Prdx1 positive group and Prdx1 negative group,VM was statistically significant difference between the two groups(?2= 12.250,P = 0.007),VM expression in Prdx1 positive group was significantly higher,the expression of P38 in Prdx1-positive patients was significantly higher than that in Prdx1-negative patients(?2= 25.593,P = 0.012),the expression of VEGF in Prdx1-positive patients was significantly higher than that in Prdx1-negative patients(?2=28.779,P=0.004).According to the clinical data of patients with lung cancer,Prdx1 expression in lymph node metastasis was significantly higher than that in patients with lymph node metastasis(?2= 20.870,P=0.000),(P <0.05).The survival time of Prdx1 positive group was shorter than that of Prdx1 negative group(P = 0.037),but other resons was no significant difference in Prdx1 positive group(P >0.05).And also the survival time of VM positive group was shorter than that of the VM negative group,the difference was statistically significant(P = 0.016).2?The expression of Prdx1 in A549 cells was significantly up-regulated by Prdx1 up-regulated plasmid and the expression of Prdx1 in H1299 cells was down-regulated by Prdx1 down-regulated plasmid,Western blot was used to identify the Prdx1 transfection efficiency.3?Wound healing and migration ability of A549 cells was significantly increased(P <0.05).And in H1299 cells it was significantly decreased after Prdx1 expression(P <0.05).4?The results of three-dimensional culture showed that the ability of A549 cells to form tubular structures was significantly increased after Prdx1 expression and H1299 cells could not form tubular structures after Prdx1 expression(P <0.05).5 ? Western blot showed the expression of E-cadherin in A549 cells was decreased and the expression of Vimentin was increased in Prdx1 cells.In H1299 cells Western blot showed the expression of E-cadherin in H1299 cells was increased and the expression of Vimentin was decreased.6?Western blotting showed that the expression of VE-cadherin and VEGF in A549 cells was significantly increased after Prdx1 up-regulation.And the expression of VE-cadherin and VEGF in H1299 cells was significantly decreased after Prdx1 down-regulation.7?Western blot showed that the expression of P38MAPK-related protein P38 remained same and P-P38 was increased in A549 cells after Prdx1 up-regulation and also showed that the expression of P38 remained same and P-P38 was decreased in H1299 cells after Prdx1 down-regulation.8?After used SB203580,Western blot showed that the expression of P-P38 and VEGF gradually decreased with the dosage of SB203580 increasing.But the expression of P38 remained same in A549 cells after Prdx1 up-regulation.Conclusion:1?Prdx1 was positively correlated with the Vasculogenic mimicry formation and lymph node metastasis of lung cancer and the survival time of Prdx1 and VM expression was shorter in positive groups,the prognosis was also poor.P38 and VEGF were highly expressed in lung cancer and correlated with Prdx1 expression2?Prdx1 can promote the migration,invasion and tubular structure of lung cancer cells,which suggesting that Prdx1 may be through the P38 MAPK pathway to affect the formation of VM in lung cancer.This study is expected to provide new therapeutic strategies for anti-vascular therapy in lung cancer patients. |
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