The Study Of Raptor Promoted Invasion And Metastasis Of Breast Cancer Cells Through The Signaling Pathway Wnt3a/?-catenin

Objective:The aim is to detect the expression of Raptor in breast cancer cells in the research and then study the role of Raptor in breast cancer cells and the molecular mechanism of invasion and metastasis through increase or reduce the expression of Raptor in the MDA231 and MCF-7,the study is to explore whether Raptor can promote the invasion and metastasis of breast cancer cells through Wnt3a/?-catenin signaling pathway.Methods:First,using Western blot to detect the protein expression of Raptor in low metastatic cell line MCF-7 and high metastatic cell line MDA231.Using siRNA technology put small RNA interference plasmid transfect into MDA231 breast cancer cells transiently to reduce the expression of Raptor in MDA231 breast cancer cells,then divided them into three groups:(1)Parental group(MDA231 group):Normal culture,no treatment,(2)Control group(Scr/MDA231 group):Transfect the control interfering plasmid,(3)Objective group(SiRaptor/MDA231 group):Transfect the interfering plasmid.Overexpression of plasmid was transfected into MCF-7 breast cancer cells transiently to increase the expression of Raptor in MCF-7 breast cancer cells,then divided them into three groups:(1)Parental group(MCF-7 group):Normal culture,no treatment,(2)Control group(MCF-7/Con group):Transfect the empty plasmid,(3)Objective group(MCF-7/Raptor group):Transfect the overexpression plasmid.Transwell and chemotaxis assay were used to detect the invasion and chemotaxis ablity of breast cancer cells.Western blot detected the expression of EMT markers;karyoplasm separation experiment tested the expression of ?-catenin in nuclear of respective group of breast cancer cellsResults:(1)Using Western blot detect the expression of Raptor protein in MDA231 and MCF-7,the result showed that the expression of Raptor was high in the MDA231,which was the high metastatic cell lines and the expression of Raptor was low in the MCF-7,which was low metastatic cell lines.Compared with the control group of Scr/MDA231 cells,the expression of Raptor protein in SiRaptor/MDA231 was decreased obviously after plasmid transfection interference.Compared with MCF-7/Con,the expression of Raptor protein was significantly increased in MCF-7/Raptor.(2)MTT experiment results showed that the influence of Raptor on cell proliferation in the groups of breast cancer cells has no statistical significance.(3)The number of cells migrated through the Transwell artificial basilemma in SiRaptor/MDA231 cells was significantly reduced than Scr/MDA231(P<0.05),and compared with MCF-7/Con,the number of migrated cells through the Transwell artificial basilemma was significantly increased in MCF-7/Raptor(P<0.05).(4)Transwell invasion and chemotaxis assay results showed that the invasion and chemotaxis ability of SiRaptor/MDA231 was decreased with the stimulation of Wnt3a,the chemotaxis ability was strongest when the concentration of Wnt3a was 100 ?g/L.(P<0.05),however,the capability of invasion and chemotaxis of MCF-7/Raptor was significantly enhanced with the stimulation of Wnt3a,the chemotaxis ability was strongest when the concentration of Wnt3a was 100 ?g/L(P<0.05)·(5)Compared with Scr/MDA231 group,the expression of Vimentin protein was decreased obviously and the expression of E-cadherin protein was increased in SiRaptor/MDA231.Compared with MCF-7/Con,the expression of Vimentin protein was increased and the expression of E-cadherin protein was reduced.(6)According to the results of Western blot method with unused Wnt3a or at the same time stimulated with Wnt3a and Dkkl,the expression of E-cadherin was higher while the expression of Vimentin and ?-catenin was reduced in SiRaptor/MDA231,whereas in the MCF-7/Raptor,the expression of E-cadherin is reduced while the expression of Vimentin and p-catenin was increased.Conclusion:1 The expression of Raptor in MCF-7 which was low metastatic cell lines was lower than in the high metastatic cell lines MDA231.It was prompt that the Raptor was associated with the invasion and metastasis of breast cancer cells.2 The ability of chemotactic movement and the invasion was enhanced through increase the expression of Raptor of breast cancer cells,the expression of Vimentin was increased and the expression of E-cadherin was reduced.The ability of chemotactic movement and the invasion was decreased through reduced the expression of Raptor of breast cancer cells,the expression of Vimentin was reduced and the expression of E-cadherin was increased.It showed that the Raptor promoted the invasion and metastasis of breast cancer cell3 The ability of chemotactic movement and the invasion was significantly enhanced,the expression of Vimentin and ?-catenin was significantly increased and the expression of E-cadherin was significantly reduced through increase the expression of Raptor of breast cancer cells with unused Wnt3a or at the same time stimulated with Wnt3a and Dkkl.The ability of chemotactic movement and the invasion was significantly decreased,the expression of Vimentin and ?-catenin was significantly reduced and the expression of E-cadherin was significantly increased through reduced the expression of Raptor of breast cancer cells with unused Wnt3a or at the same time stimulated with Wnt3a and Dkkl.So Raptor can promote the invasion and metastasis of breast cancer cells through Wnt3a/?-catenin signaling pathway.