ABSTRACT
BACKGROUND: Ubiquitin specific peptidase 39 (USP39), an essential factor in the assembly of the mature spliceosome complex, has an aberrant expression in several cancer. However, its function and the corresponding mechanism on human osteosarcoma has not been fully explored yet. METHODS: The mRNA and DNA copies of USP39 were increased in osteosarcoma cancer tissues compared with the one in human normal tissues according to datasets from the publicly available Oncomine database. A further western blot analysis also demonstrated an aberrant endogenous expression of USP39 in three different osteosarcoma cells. Then lentivirus-mediated short hairpin RNA (shRNA) was designed to silence USP39 in human osteosarcoma cell line U2OS, which is used to test the impact of USP39-silencing on cellular proliferation, colony formation, cell cycle distribution and apoptosis. RESULTS: Knockdown of USP39 expression in U2OS cell significantly decreased cell proliferation, impaired colony formation ability. A further analysis indicated suppression of USP39 arrested cell cycle progression at G2/M phase via p21 dependent way. In addition, the results of Annexin V/7-AAD staining suggested the knockdown of USP39 could promote U2OS cell apoptosis through PARP cleavage. CONCLUSIONS: These results uncover the critical role of USP39 in regulating cancer cell mitosis and indicate USP39 is critical for osteosarcoma tumorigenesis.
Subject(s)
Humans , Osteosarcoma/enzymology , Osteosarcoma/pathology , Apoptosis , Gene Knockdown Techniques/methods , Ubiquitin-Specific Proteases/metabolism , Tumor Stem Cell Assay , Gene Expression Regulation, Neoplastic , Lentivirus , Cell Line, Tumor , Cell Proliferation , Ubiquitin-Specific Proteases/genetics , Flow Cytometry , Genetic VectorsABSTRACT
Matrixmetalloproteinases [MMPs] are a class of matrix basement membrane degrading enzymes which were shown to be associated with metastases in several human tumors. This study is aimed to investigate the potential effect of MMP-9 in imparting the frequently reported different behavioral pattern between jaws and long bones osteosarcomas. Tissue blocks of ten cases of jaws osteosarcomas and another ten of long bones osteosarcomas were collected and stained immunohistochemically with monoclonal antibodies to MMP-9. The majority of cases [70%] were positive for MMP-9 expression which indicates a role in tumor spread, however; there was no significant difference between the sites. The study indicates that MMP-9 is probably not involved in the biologic differences between jaws and long bones osteosarcomas
Subject(s)
Humans , Male , Female , Adult , Osteosarcoma/enzymology , Osteosarcoma/pathology , Immunohistochemistry , Jaw/pathology , Retrospective StudiesABSTRACT
Osteosarcoma is the most common primary bone tumor, but the pathogenesis is not well understood. While cyclooxygeanse-2 (COX-2) is known to be closely associated with tumor growth and metastasis in several kinds of human tumors, the function of COX-2 in osteosarcoma is unclear. Therefore, to investigate the function of COX-2 in osteosarcoma, we established stable cell lines overexpressing COX-2 in U2OS human osteosarcoma cells. COX-2 overexpression as well as prostaglandin E(2) treatment promoted proliferation of U2OS cells. In addition, COX-2 overexpression enhanced mobility and invasiveness of U2OS cells, which was accompanied by increases of matrix metalloproteinase-2 and -9 (MMP-2 and -9) activities. Selective COX-2 inhibitors, NS-398 and celecoxib, inhibited cell proliferation and abrogated the enhanced mobility, invasiveness and MMP activities induced by COX-2 overexpression. These results suggest that COX-2 is directly associated with cell proliferation, migration and invasion in human osteosarcoma cells, and the therapeutic value of COX-2 inhibitors should be evaluated continuously.