NEDD9 Promotes Invasion And Migration Of Colorectal Cancer Via JNK/EMT

Objective:Colorectal cancer is a common malignant tumor,the second leading cause of cancer death.The occurrence of invasion and metastasis of colorectal cancer is the main cause of low survival rate and high mortality.The causes and mechanisms of invasion and metastasis of colorectal cancer are still not clear,so it is particularly important to study molecular markers that cause invasion and metastasis of colorectal cancer.Neural precursor cell expressed developmentally down-regulated 9(NEDD9,also known as HEF1 and Cas-L),which is a multi-domain skeletal protein that is highly expressed in various tumors of melanoma,liver cancer,lung cancer and breast cancer.The purpose of this study was to investigate the expression of NEDD9 in colorectal cancer tissues and cells,and NEDD9 through stress-activated protein kinase(JNK,c-Jun N-terminal kinase)/ epithelial mesenchymal transition(EMT)signaling pathway The effect on invasion and metastasis of colorectal cancer.While exploring the pathogenesis of colorectal cancer,it provides targets for its diagnosis and cancer drug treatment.Methods:The expression of NEDD9 in 40 colorectal cancer tissues and adjacent normal tissues,as well as in normal colorectal cells FHC and colorectal cancer cells HCT-116 were detected by quantitative PCR.The experiment was divided into two groups: Lv-NC and Lv-NEDD9.HCT-116 cells were transfected with blank vector lentivirus as Lv-NC group,while HCT-116 cells were transfected with NEDD9 down-regulated lentivirus as LvNEDD9 group.The expressions of NEDD9,EMT related markers protein E-cadherin and Vimentin in the two groups were detected by quantitative PCR.Transwell invasion assay was used to detect the invasive ability of the two groups of cells,and the migration ability of the two groups was detected by Transwell migration assay and scratch migration assay.The proliferation ability of the two groups of cells was examined by CCK-8 assay.Western Blot assay was used to detect the expression levels of NEDD9,JNK,E-cadherin and Vimentin proteins in the two groups.After adding JNK blocker,the experiment was divided into two groups: DMSO and SP600125.DMSO was added to HCT-116 cells as DMSO group,and SP600125 was added to HCT-116 cells as SP600125 group.Transwell invasion assay was used to detect the invasive ability of the two groups of cells,and the migration ability of the two groups was detected by Transwell migration assay and scratch migration assay.The proliferation ability of the two groups of cells was examined by CCK-8 assay.Western Blot assay was used to detect the expression levels of JNK,E-cadherin and Vimentin proteins in the two groups of cells.Results:Quantitative PCR results showed that NEDD9 was highly expressed in colorectal cancer tissues compared with adjacent tissues,statistical significance(P<0.01).Compared with normal colorectal cells,NEDD9 was highly expressed in colorectal cancer cells,statistical significance(P<0.01).Compared with the Lv-NC group,NEDD9 was successfully down-regulated in the Lv-NEDD9 group,the expression of Vimentin was decreased,and the expression of E-cadherin was increased,which was statistically significant(P<0.01).Transwell results showed that compared with the Lv-NC group,the migration(P<0.05)and invasion(P<0.01)ability of the Lv-NEDD9 group was weakened.Compared with the DMSO group,the migration and invasion ability of the SP600125 group was weakened(P<0.01).The results of the wound healing assay showed that the migration ability of the Lv-NEDD9 group was weaker than that of the Lv-NC group(P<0.01).Compared with the DMSO group,the migration ability of the SP600125 group was weakened(P<0.01).Western Blot results showed that compared with Lv-NC group,the expression of NEDD9,JNK and Vimentin in Lv-NEDD9 group decreased(P<0.05),and the expression of E-cadherin increased(P<0.01).Compared with DMSO group,the expression of JNK and Vimentin in SP600125 group decreased(P < 0.01),and the expression of E-cadherin increased(P < 0.05).Conclusion:NEDD9 is highly expressed in colorectal cancer tissues and cells,and NEDD9 can promote the invasion and migration of colorectal cancer through JNK/EMT signaling pathway.