The Biological Functions And Mechanism Of Tumor Suppressor Gene ADAMTS9 In Colorectal Cancer

Objective:Our aim is to detect that the expression of tumor-suppressor gene(TSG)ADAMTS9 in many species of colorectal cancer cells and 32 pairs of clinical colorectal cancer samples. Meanwhile, we aim to investigate the function of ADAMTS9 and the underlying mechanism in colorectal cancer in vitro.Methods:1, We used RT-PCR and real-time PCR to detect the expression of ADAMTS9 in colorectal cancer cell lines, colorectal cancer tissues and adjacent cancer tissues.2, We transfected ADAMTS9 into colorectal cancer cell lines CaCo2 and HCT116 to analysis the influence of overexpression ADAMTS9 on colorectal cancer cells apoptosis, cycle, growth, proliferation and migration by flow cytometry instrument technology, cloning experiment, CCK-8assay, and transwell migration experiment. In addition, We transfected shRNA-ADAMTS9 into colorectal cancer cell line SW480 to analysis theinfluence of down-regulated expression ADAMTS9 on colorectal cancer cells proliferation by CCK8 assay.3, We analyzed the effect of ADAMTS9 on AKT signaling pathways by western blot experiment to demonstrate the possible mechanisms of ADAMTS9 affecting the formation of colorectal cancer.Results:1, ADAMTS9 was downregulated or silenced in 83.3%(5/6) of CRC cell lines and in 68.75%(22/32)of CRC tissues(p<0.05).2, Overexpression of ADAMTS9 in the cell lines CaCo2 and HCT116 can significantly promote its apoptosis and inhibit cells proliferation and migration, flow cytometer analysis showed that it can inhibit colorectal cancer cells grow in G0-G1 phase. On the other hand, shRNA mediated knockdown of ADAMTS9 in cell line sw480 can significantly promote cell growth.3, Western blot experiment showed ADAMTS9 had an effect on colorectal cancer biology functions by influencing the AKT signaling pathway.Conclusion:Tumor-suppressor gene ADAMTS9 was frenquently down-regulated or silenced in CRC and had a significant inhibition effect on colorectalcancer. which may be a promissing epigenetic markers for early diagnosis and prognosis evaluation of colorectal cancer.