Effect Of Nodal On Hypoxia-induced Invasion And Migration In Malignant Melanoma A375 Cells

Objective:To discuss the influence of Nodal signaling pathway in-duced by hypoxia on malignant melanoma A375 cells invasion and migration.Methods:The chemicals Cocl₂ of different concentration based on different time of exposure were used to induce the production of A375 cells in hypoxia micro-environment.To confirm drug's appropriate concentration and time of exposure by detecting cell proliferation viability by MTT assay.The expression of HIF-1?was detected by Western Blot,as well as the expression level of Nodal and its receptor ALK4/7.In addition,the influence on expression of protein E-cadherin,MMP2 and Vimentin which related to the invasion and migration were detected by Western Blot in melanoma.Meanwhile,the protein level of LDH-A and LDH-B which related to glucose metabolism were also detected in hypoxia.Nodal signal activation induced by hypoxia environment in melanoma were interfered by the small molecule inhibitor SB-431542 with appropriate concentration and time of exposure.Results:The hypoxia micro-environment was successfully induced by Cocl₂ on concentration of 150?mol/L effective for 48 hours in A375 cells.The expression of Nodal and receptor ALK7 were induced by hypoxia?P<0.05?,and the up-regulation of Nodal signal in cells was productively inhibited by SB-431542 on concentration of10?mol/L effective 48 hours.The protein levels of migration and invasion markers such as MMP2 and Vimentin were upregulated by Nodal in hypoxia?P<0.05?.Meanwhile,Hypoxia-induced Nodal could also lead to the increase of LDH-A expression and decrease expression of LDH-B protein level?P<0.05?.All of the above changes were partially reversed by SB-431542?P<0.05?.Conclusion:The hypoxia environment can promote the malignant processes of migration and invasion in A375 cells,and the cells tend to be energy supplied through glycolysis.It may be related to the activation of Nodal signal induced by HIF-1?.