| Objective:Laryngeal Squamous Cell Carcinoma(LSCC)has a high incidence in malignant tumors of the head and neck.In spite of recent advances in clinical studies,laryngeal squamous cell carcinoma(LSCC)has no significant improvement in survival rates compared with the past.Golgi membrane protein1(GOLM1)is abnormally expressed in a variety of human malignant tumors,but the biological behavior of GOLM1 in LSCC is not clear at present.mi R-27 b is a common mi RNA with abnormal regulation in human cancer.Through the inhibition of multiple targets,mi R-27 b has recently become a key inhibitor or oncogene in cancer.The purpose of this study was to investigate the expression of Golgi membrane protein GOLM1 in laryngeal squamous cell carcinoma(LSCC)and its effect on the proliferation,migration and invasion of LSCC cells.To explore the relationship between GOLM1 and Wnt/β-catenin pathway in LSCC,and to verify its possible upstream regulatory mechanism.In order to provide experimental basis for LSCC to find new tumor markers and molecular targets for LSCC.Methods: Part I.The effect of GOLM1 on laryngeal squamous cell carcinoma through Wnt/β-catenin pathway: 1.The expression of GOLM1 in LSCC and paracancer tissues was detected by immunohistochemistry.2.Synthesis of si RNA,overexpression vector and negative control for GOLM1.3.The effect of GOLM1 on the proliferation of LSCC cells was detected by CCK8.4.The influence of GOLM1 on the migration ability of LSCC was detected by scratch experiment.5.Transwell assay was used to detect the effect of GOLM1 on the invasion ability of LSCC cells.6.The effect of GOLM1 on apoptosis and cell cycle distribution of LSCC cells was detected by flow cytometry.7.The effect of GOLM1 on Wnt/β-catenin pathway in LSCC was detected by q RT-PCR and Western blot.Part Ⅱ:The mechanism of mi R-27b-3p targeting GOLM1 affecting laryngeal LSCC: 1.The expression of mi R-27b-3p in LSCC and adjacent tissues was detected by q RT-PCR.2.Dual luciferase was used to detect the targeting relationship between mi R-27b-3p and GOLM1.3.q RT-PCR and Western blot were used to detect the effects of mi R-27b-3p on GOLM1 and Wnt/β-catenin pathways in LSCC.4.CCK8 method verified the effect of mi R-27b-3p on the proliferation ability of LSCC cells through GOLM1.5.Scratch test verified the influence of mi R-27b-3p on LSCC migration ability through GOLM1.6.Transwell experiment verified the effect of mi R-27b-3p on the invasion ability of LSCC cells through GOLM1.7.Flow cytometry was used to detect the effects of mi R-27b-3p on apoptosis and cell cycle distribution of LSCC cells through GOLM1.8.The laryngeal carcinoma tumor model in nude mice was established to verify the influence of mi R-27b-3p/GOLM1 axis on LSCC at the animal level.Result: 1.GOLM1 promotes LSCC progression through the Wnt/β-catenin pathway1.1.GOLM1 was significantly overexpressed in LSCC.1.2.GOLM1 promoted the proliferation,migration and invasion of LSCC cells,inhibited cell apoptosis,and promoted cells to enter the S phase.1.3.GOLM1 positively regulates the Wnt/β-catenin pathway in LSCC.2.mi R-27b-3p targeting GOLM1 affects the biological behavior of LSCC cells: 2.1.Low expression of mi R-27b-3p in LSCC.2.2.mi R-27b-3p inhibited the proliferation,migration and invasion of LSCC cells,promoted cell apoptosis,and inhibited cells from entering the S phase.2.3.mi R-27b-3p inhibits GOLM1 expression and Wnt/β-catenin pathway in LSCC.2.4.mi R-27b-3p inhibited tumor growth and inhibited the expression of GOLM1 and Wnt/β-catenin pathway in tumor models of nude mice.Conclusion: GOLM1 is highly expressed in LSCC and promotes the proliferation,migration and invasion of LSCC cells through the Wnt/β-catenin pathway.This process is regulated by the targeted mi R-27b-3p,which can inhibit the effects of GOLM1/Wnt axis on the proliferation,migration and invasion of LSCC cells as well as the cell cycle. |
