Effect And Mechanism Of Activin A On Apoptosis And Migration Of Mouse Myeloma NS-1 Cells

Activin A belongs to transforming growth factor β(TGF-β)Superfamily of multifunctional immunoregulatory factors,with a wide range of histological distribution.Activin A plays an important role in the development of many kinds of tumors.However,it is still unclear whether activin A can be expressed in mouse myeloma NS-1 cells,and whether exogenous activin A can regulate apoptosis and migration of mouse myeloma NS-1 cells through Smad3 signaling.Therefore,this study explored the effect of activin A on apoptosis and migration of mouse myeloma NS-1 cells by culturing them in vitro,the effect of activin A on apoptosis of mouse myeloma NS-1 cells was verified by overexpression of Smad3 and knockdown of Smad3,which further revealed the mechanism of activin a regulating biological activity of mouse myeloma cells.1.Methods(1)Flow cytometry was used to detect the apoptosis of NS-1 cells with double staining of Annexin V and 7-AAD.(2)Transwell chamber method was used to detect the migration of NS-1 cells.(3)RT-PCR was used to detect the expression of signal molecules and migration related genes.(4)Western blotting was used to detect the expression of migration related molecular proteins.(5)pc DNA3-Smad3 and Smad3 sh RNA expression plasmids were transfected into NS-1 cells for Smad3 overexpression and knockdown.2.Results(1)Activin βA,Smad signal proteins and Activin receptors were expressed in mouse myeloma NS-1 cells.(2)Activin A inhibited the proliferation and promoted apoptosis of NS-1 cells.(3)Transwell chamber assay showed that activin A inhibited the migration of mouse myeloma NS-1 cells,RT-PCR and Western blotting showed that activin A inhibited the expression of MMP2 and MMP9,and up-regulated the expression of TIMP1 and TIMP2,Flow cytometry showed that activin A inhibited Ca2 + influx in NS-1 cells.(4)pc DNA3-smad3 and Smad3 sh RNA expression plasmids were transfected into mouse myeloma NS-1 cells,respectively.The results showed that the apoptotic rate of NS-1 cells in Smad3 knockdown group was significantly lower than that of the empty plasmid group,and Smad3 knockdown could inhibit the effect of Activin A on promoting apoptosis of NS-1 cells.On the contrary,the apoptotic rate of NS-1 cells in Smad3 overexpression group was significantly higher than that of the empty plasmid group.The apoptotic rate of NS-1 cells in Smad3 overexpression group was more obvious treated with Activin A.3.ConclusionIn conclusion,Activin A can inhibit the proliferation and promote the apoptosis of mouse myeloma NS-1 cells,which is related to the up regulation of Smad3 signaling pathway by Activin A.In addition,Activin A can inhibit the migration of mouse myeloma NS-1 cells,which is related to the decrease expression of MMP2 and MMP9 and the inhibition of calcium influx.