Progression of human osteosarcoma with altered ANT1/SLC25a4 expression.

Abstract

Introduction: Osteosarcoma (OS) is the most prevalent primary bone malignancy in children and adolescents. Bioinformatics analyses have identified ANT1 (encoded by SLC25A4) as part of a gene cluster implicated in the development and progression of osteosarcoma. Authors of this study investigated the expression of ANT1/SLC25A4 in clinical specimens and established OS cell lines to evaluate its potential as a prognostic biomarker and its mechanistic role in OS pathogenesis.

Methods: Immunohistochemical (IHC) analysis of ANT1 expression was performed on 63 archived human OS tissue sections (IRB-exempt). To assess functional impact, commercial MG-63 and HOS osteosarcoma cell lines were genetically modified to achieve SLC25A4 overexpression or knockdown. Cellular behavior was evaluated using proliferation, scratch wound-healing, and transwell invasion assays. In vitro numerical data were analyzed using one-way ANOVA with the LSD method for post hoc multiple comparisons. Correlations with clinical prognostic indicators were assessed using bivariate correlation analysis (SPSS v.22).

Results: IHC staining revealed that most clinical OS specimens (48/63) exhibited significantly diminished ANT1 expression compared with normal periosteal tissue. In vitro, SLC25A4 knockdown significantly accelerated cell proliferation. Conversely, SLC25A4 overexpression significantly inhibited cell migration and invasion (p <0.05) compared with controls.

Conclusions: Our findings demonstrate that ANT1/SLC25A4 frequently is downregulated in clinical OS, and that its restoration suppresses aggressive cellular phenotypes in vitro. These results suggest that SLC25A4 may function as a tumor suppressor in osteosarcoma. Further research is needed to correlate SLC25A4 expression with patient survival and to define the underlying molecular pathways.