Abstract
BackgroundOsteosarcoma is a malignant bone tumor. Increasing evidences have revealed that a disintegrin and metalloproteinase 10 (ADAM10) is implicated in tumor development. The main purpose of this study is to explore the effects of ADAM10 on osteosarcoma cell functions and the underlying molecular mechanisms.MethodsWestern blot and quantitative real-time PCR were performed to detect the expression of ADAM10 in one osteoblast (hFOB 1.19) and six osteosarcoma cells (Saos-2, SW1353, HOS, U-2OS, MG63, and 143B). The biological functions of ADAM10 in osteosarcoma cells were measured by cell counting kit-8 assay, flow cytometry, wound healing assay, and transwell assay. The interaction between miR-122-5p and ADAM10 was validated using dual-luciferase reporter assay. The effect of ADAM10 on the tumorigenicity of osteosarcoma cells was evaluated in a nude mice model in vivo.ResultsWe found that the expression of ADAM10 was relatively high in osteosarcoma cells compared with that in osteoblast. ADAM10 promoted osteosarcoma cell growth, migration, and invasion. Mechanism studies showed that knockdown of ADAM10 inactivated E-cadherin/β-catenin signaling pathway, as evidenced by increased the level of E-cadherin, reduced nuclear translocation of β-catenin, and decreased the levels of MMP-9, Cyclin D1, c-Myc, and Survivin. Downregulation of ADAM10 suppressed the tumorigenicity of osteosarcoma cells in vivo. Furthermore, ADAM10 was validated to be a downstream target of microRNA-122-5p (miR-122-5p). MiR-122-5p-induced inhibition of cell proliferation, migration, and invasion was reversed by overexpression of ADAM10 in osteosarcoma cells.ConclusionsCollectively, the key findings of this study are that ADAM10 promotes osteosarcoma cell proliferation, migration, and invasion by regulating E-cadherin/β-catenin signaling pathway, and miR-122-5p can target ADAM10, indicating that miR-122-5p/ADAM10 axis might serve as a therapeutic target of osteosarcoma.
Highlights
a disintegrin and metalloproteinase 10 (ADAM10) expressions were higher in the osteosarcoma cells We investigated the ADAM10 expression in one osteoblast and six osteosarcoma cells (Saos-2, SW1353, HOS, U-2OS, MG63, and 143B) through western blot and real-time PCR, ADAM10 knockdown decreased osteosarcoma cell proliferation, migration and invasion but increased cell apoptosis the U-2OS and MG63 cells with higher ADAM10 expressions were adopted to do the transfection with two ADAM10 shRNAs to knock down its expressions
ADAM10 knockdown affected E‐cadherin/β‐catenin signaling pathway in the osteosarcoma cells In order to investigate the effects of ADAM10 knockdown on E-cadherin/β-catenin signaling pathway, the U-2OS and MG63 cells were transfected with ADAM10shRNA for 48 h
The decreased levels of soluble E-cadherin were found in supernatants of ADAM10-silenced U-2OS and MG63 cells as measured by ELISA with a soluble E-cadherin–specific antibody (Fig. 4b)
Summary
The main purpose of this study is to explore the effects of ADAM10 on osteosarcoma cell functions and the underlying molecular mechanisms. ADAM family plays an important role in cell proliferation, migration, invasion, and angiogenesis, and it is related to the metastasis of human tumors [3]. The previous studies revealed that the expression of ADAM10 was overexpressed in a variety of tumors including liver cancer [5], melanoma [6], gastric cancer [7], lung cancer [8], pancreatic cancer [9], and bladder cancer [10]. To the best of our known, the effects of ADAM10 on cell functions and the underlying molecular mechanisms in osteosarcoma have never been reported
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