The Role And Mechanism Of ADAM23in Proliferation、Invasion And Migration In Lung Squamous Cell Carcinoma SK-MES-1Cells

Background and Objective:Lung cancer is one of the most serious malignant tumors in developed country and the whole world, since then, invasion and metastasis have become the major factors to death of lung cancer patients. Our previous studies showed that the decrease expression of ADAM23was related with metastasis and malignant progression in non-small cell lung, but the molecular mechanism is still unknown.A disintegrin and metalloprotease23belongs to ADAM family, which contain disintegrin domain, and sharing metalloproteinase domain with matrix metalloproteinase. The member of the family are known to determine cell-cell adhesion、cell migration and proliferation and other biological behaviors, simultaneously, as a tumor suppressor gene, the deficiency of ADAM23is directly involved in the processes of tumors.The epithelial-to-mesenchymal transition is an important process of embryonic development, but it can be abnormally activated in malignant tumors which is the early events in tumor invasion and metastasis. Early experiments showed that the decrease expression of ADAM23contribute to the increase expression of αvβ3, which play a collaboration role in the invasion and metastasis in non-small lung cancer.Many signaling pathways are involved in invasion and metastasis, we focus our attention to PI3K/AKT since it can be activated by ay which is the member of integrin family. There have been reported that the consistent activation of PI3K/AKT signaling pathway cay promote the process of EMT. AKT is the central of this signaling pathway, the activation of AKT plays important role in cell growth, proliferation, motility and cell invasion.Our findings demonstrate that the decrease expression of ADAM23can promote the ability of proliferation、invasion and migration in SK-MES-1cells, as well as the pAKT expression level. While, under low expression of ADAM23, the inhibition of the PI3K/AKT pathway have changed the invasion and migration ability. This study is investigate the role and mechanism of ADAM23in proliferation、invasion and migration in lung squamous cell SK-MES-1.Methods:(1) Using the liposome-mediated transfection techniques, transient1y transfect ADAM23-siRNA-1,2,3and negative control into lung squamous cell SK-MES-1, using untreated SK-MES-1cell as blank control, detecting the relative expression of ADAM23mRNA, selected the most inhibit efficiency of ADAM23-siRNA;(2)Transient transfection ADAM23-siRNA-3、negative control into SK-MES-1cell, let untreated SK-MES-1cell as blank control, investigate the protein expression of TWSIT、E-cadhern、α-SMA、vimentin;(3)Investigate the ability of cell proliferation of ADAM23-siRNA treated cells in MTT assay;(4)To detect the ability of cell invasion and migration of ADAM23-siRNA treated cells by transwell assay;(5)To detect the expression of AKT, pAKT, av in ADAM23-siRNA treated cells by western blot;(6) Treat ADAM23-siRNA-3cells as study object, added LY29402、DMSO, detect the expression of AKT、pAKT、αV of the LY294002treated cells by western blot;(7)To detect the expression of TWSIT、E-cadherin、 α-SMA、vimentin after treated by specific inhibition of PI3K/AKT;(8)The ability of invasion and migration of cells after treated by LY294002.Results:(1)All of the siRNA can availably inhibit the expression of ADAM23mRNA, but ADAM23-siRNA-3is the best one;(2)Compared with the blank and negative control, the expression of E-cadherin after ADAM23-siRNA treated was decreased, while the expression of a-SMA, vimentin, TWSIT were increased;(3)The ability of cell proliferation of ADAM3-siRNA cells was significantly stronger than negative and blank control(P<0.05);(4)In vitro assay, ADAM23-siRNA treated group showed higher ability of invasion and migration (P<0.05);(5)Compared with the blank and negative control, the expression of pAKT, av after ADAM23-siRNA treated was increased, while the there is no effect to AKT;(6)When treated with the specific inhibition of PI3K/AKT, the expression of av after ADAM23-siRNA treated was decreased, pAKT was missing, but have no effect to AKT;(7) After the treatment of LY294002, the expression of E-cadherin was downregulated, while α-SMA、vimentin、TWSIT were upregulated;(8)After the treatment of LY294002, the ability of cell invasion and migration was downregulated(P<0.05).Conclusion:(1). The silencing of ADAM23can promote the ability of proliferation, invasion and metastasis in lung squamous cell carcinoma SK-MES-1.(2) PI3K/AKT signaling pathway may involved in the promotion of invasion and metastasis ability in lung squamous cell carcinoma resulted from the downregulation of ADAM23.(3) The interaction between ADAM23and integrin av may activate PI3K/AKT signaling pathway and modulates invasion and metastasis in lung squamous cell carcinoma.