Qing Wang1, Ming-Jiang Liu2, Jie Bu3, Jian-Liang Deng2, Bin-Yuan Jiang4, Liang-Dong Jiang5, Xiao-Jie He6. 1. Department of Respiratory and Critical Care Medicine, Changsha Central Hospital (University of South China Affiliated Changsha Central Hospital), Changsha 410004, Hunan Province, PR China. 2. Department of Orthopaedics, Changsha Central Hospital (University of South China Affiliated Changsha Central Hospital), Changsha 410004, Hunan Province, PR China. 3. Orthopedics & Soft Tissue Department, Hunan Cancer Hospital, Changsha 410013, Hunan Province, PR China. 4. Central Laboratory, Changsha Central Hospital (University of South China Affiliated Changsha Central Hospital), Changsha 410004, Hunan Province, PR China. 5. Department of Orthopaedics, Changsha Central Hospital (University of South China Affiliated Changsha Central Hospital), Changsha 410004, Hunan Province, PR China. Electronic address: 806602491@qq.com. 6. Laboratory of Pediatric Nephrology, Institute of Pediatrics, Central South University, Changsha 410011, Hunan Province, PR China. Electronic address: xiaohewe542@163.com.
Abstract
AIMS: Osteosarcoma (OS) is an extremely malignant bone cancer with high incidence and rapid progression. This study aims to investigate the role and underlying mechanisms of MALAT1 and miR-485-3p in OS. MATERIALS AND METHODS: qRT-PCR and Western blotting were utilized to measure the levels of miR-485-3p, MALAT1, c-MET, AKT3, p-mTOR, mTOR, glycolysis-related proteins or migration-related proteins. Colony formation and transwell assay were used to test the roles of miR-485-3p, MALAT1, c-MET and AKT3 in cancer cell proliferation, migration and invasion. Dual luciferase assay was used to validate the interactions of miR-485-3p/c-MET, miR-485-3p/AKT3, and MALAT1/miR-485-3p. Glucose uptake assay and measurement of lactate production were employed to determine the glycolysis process. Mouse tumour xenograft model was used to determine the effect of shMALAT1 and miR-485-3p mimics on tumour growth and metastasis in vivo. KEY FINDINGS: miR-485-3p was decreased while c-MET, AKT3, and MALAT1 were increased in human OS tissues and cells. miR-485-3p bound directly to c-MET and AKT3 mRNAs and repressed OS cell glycolysis, proliferation, migration, and invasion through decreasing glycolysis-related proteins and migration-related proteins via inhibiting c-MET and AKT3/mTOR pathway. In addition, MALAT1 interacted with miR-485-3p and disinhibited c-MET and AKT3/mTOR signalling. Knockdown MALAT1 or overexpression of miR-485-3p restrained OS tumour growth and lung metastasis in vivo. SIGNIFICANCE: miR-485-3p suppresses OS glycolysis, proliferation, and metastasis via inhibiting c-MET and AKT3/mTOR signalling and MALAT1 acts as a sponge of miR-485-3p. MALAT1 and miR-485-3p may be the key regulators in OS progression, and potential molecular targets for future OS therapy.
AIMS: Osteosarcoma (OS) is an extremely malignant bone cancer with high incidence and rapid progression. This study aims to investigate the role and underlying mechanisms of MALAT1 and miR-485-3p in OS. MATERIALS AND METHODS: qRT-PCR and Western blotting were utilized to measure the levels of miR-485-3p, MALAT1, c-MET, AKT3, p-mTOR, mTOR, glycolysis-related proteins or migration-related proteins. Colony formation and transwell assay were used to test the roles of miR-485-3p, MALAT1, c-MET and AKT3 in cancer cell proliferation, migration and invasion. Dual luciferase assay was used to validate the interactions of miR-485-3p/c-MET, miR-485-3p/AKT3, and MALAT1/miR-485-3p. Glucose uptake assay and measurement of lactate production were employed to determine the glycolysis process. Mousetumour xenograft model was used to determine the effect of shMALAT1 and miR-485-3p mimics on tumour growth and metastasis in vivo. KEY FINDINGS:miR-485-3p was decreased while c-MET, AKT3, and MALAT1 were increased in human OS tissues and cells. miR-485-3p bound directly to c-MET and AKT3 mRNAs and repressed OS cell glycolysis, proliferation, migration, and invasion through decreasing glycolysis-related proteins and migration-related proteins via inhibiting c-MET and AKT3/mTOR pathway. In addition, MALAT1 interacted with miR-485-3p and disinhibited c-MET and AKT3/mTOR signalling. Knockdown MALAT1 or overexpression of miR-485-3p restrained OS tumour growth and lung metastasis in vivo. SIGNIFICANCE: miR-485-3p suppresses OS glycolysis, proliferation, and metastasis via inhibiting c-MET and AKT3/mTOR signalling and MALAT1 acts as a sponge of miR-485-3p. MALAT1 and miR-485-3p may be the key regulators in OS progression, and potential molecular targets for future OS therapy.
Authors: Lauretta Levati; Cristian Bassi; Simona Mastroeni; Laura Lupini; Gian Carlo Antonini Cappellini; Laura Bonmassar; Ester Alvino; Simona Caporali; Pedro Miguel Lacal; Maria Grazia Narducci; Ivan Molineris; Federica De Galitiis; Massimo Negrini; Giandomenico Russo; Stefania D'Atri Journal: Cancers (Basel) Date: 2022-07-29 Impact factor: 6.575