Evaluation of local and circulating osteopontin in malignant and benign primary bone tumors

J Bone Oncol. 2021 Jun 19:29:100377. doi: 10.1016/j.jbo.2021.100377. eCollection 2021 Aug.

Abstract

Purpose: The development of novel and efficient biomarkers for primary bone cancers is of grave importance.

Methods: The expression pattern of osteopontin (OPN) was investigated in the 153 patients with benign (n = 72) and malignant (n = 81) primary bone cancers. Both local and circulating OPN mRNA expression levels and their protein concentration in serum and tumor site were assessed using real-time qRT-PCR, ELISA, and immunohistochemistry techniques, respectively. As a control, 29 healthy individuals were considered. The number of 153 tumor tissue specimens and the 153 paired margins were taken on surgical resection from the patients. 153 blood samples were also drained from all participants, then peripheral blood mononuclear cells (PBMC) and sera were separated.

Results: The mean mRNA expression was significantly higher in all of the cancerous tissues than the paired margins and the PBMC of the patients than the controls. Consistently, the protein concentrations of OPN in serum and tumor tissues were significantly higher in the patients. Furthermore, the malignant cases had significantly elevated the mRNA levels and the protein compared to the benign cases. OPN could potentially differentiate the patients from the controls with 100% sensitivity and specificity in serum. Moreover, OPN could predict some of the malignant cases' clinicopathological features, including metastasis, recurrence, grade, and response to chemotherapy.

Conclusions: In conclusion, OPN might be involved in the pathogenesis of primary bone tumors and can be considered as a potential biomarker to bone cancer diagnosis.

Keywords: ANOVA, One-way analysis of variance; AUC, area under the curve; Bone tumors; CI, confidence interval; Chondrosarcoma; DAPI, 4′,6-Diamidine-2′-phenylindole dihydrochloride; ELISA, Enzyme-linked immunosorbent assay; EMT, epithelial-mesenchymal transition; Ewing Sarcoma; HIF-1α, hypoxia-inducible factor-1 alpha; HRP, horseradish peroxidase; MMP9, Matrix metallopeptidase 9; OCT, Optimal Cutting Temperature; OPN, Osteopontin; Osteopontin; Osteosarcoma; PBMC, Peripheral blood mononuclear cells; PBS, phosphate-buffered saline; ROC, receiver operating characteristic; S100A8, S100 calcium-binding protein A8; SOX9, SRY-Box Transcription Factor 9; cDNA, Complementary DNA; qRT-PCR, Quantitative Real-time transcription-polymerase chain reaction.