SiRNA-Ambra1 Enhances The Proliferation And Migration Of Colorectal Cancer Cells

Objective: To investigate the effect of Ambra1 in colorectal cancer SW480 and HCT15 cells,and to explore its possible mechanism.Methods: Colorectal cancer specimens and tissue adjacent to carcinoma in clinical were collected,then RNA and proteins of the specimens were extracted,then we use QPCR and WB to know the expression about Ambra1.the same method will be used in CRC cell lines and normal intestinal epithelial cell line(NCM460),siRNA-Ambra1 targeted Ambra1 gene was transfected into colorectal cancer cells,the transfection effect were detected by qPCR and WB.Then we use the routine method to check the phenotype effect in CRC cells by siRNA-Ambra1,include migration,invasion and proliferation.The effects of siRNA-Ambra1 on the expressions of epithelial-mesenchymal transition(EMT)related proteins(N-cadherin,E-cadherin and Vimentin),extracellular singal-regulated kinase(ERK)related protein and c-myc in colorectal cancer SW480 and HCT15 cells were detected by Western blotting.And build a stable low expression of Ambra1 colorectal cancer cell lines(HCT116 and KM12),animal experiments in vivo will be perform.Results: Ambra1 is high expressed not only in normal clinical tissues,but also in NCM460 cell.Ambra1 can be downed by siRNA-Ambra1 and promoted the colony formation,proliferation,migration and invasion of SW480 and HCT15 cells(all P < 0.05).siRNA-Ambra1 up-regulated theexpression level of EMT related protein N-cadherin(P < 0.05),and E-cadherin’s expression level is down(P < 0.05),up-regulated the expression level of ERK signal pathway related protein phosphorylated ERK1/2(p-ERK1/2)(P < 0.05).In vivo,knockdown Ambra1 can promote the proliferation in colorectal cancer cell(P < 0.05).Conclusions: Ambra1 as a protective factor exists in human body.Knockdown Ambra1 gene can enhance the proliferation,migration and invasion of colorectal cancer SW480 and HCT15 cells via EMT and ERK signaling pathways.