Neoadjuvant multidrug chemotherapy including high-dose methotrexate modifies VEGF expression in osteosarcoma: an immunohistochemical analysis.

Background: Angiogenesis plays a role in the progression of osteosarcoma, as well as in other mesenchymal
tumors and carcinomas, and it is most commonly assessed by vascular endothelial growth factor (VEGF) expression
or tumor CD31-positive microvessel density (MVD). Tumor VEGF expression is predictive of poor prognosis, and
chemotherapy can affect the selection of angiogenic pattern. The aim of the study was to investigate the clinical
and prognostic significance of VEGF and CD31 in osteosarcoma, both at diagnosis and after neoadjuvant
chemotherapy, in order to identify a potential role of chemotherapy in angiogenic phenotype.
Methods: A retrospective analysis was performed on 16 patients with high grade osteosarcoma. In each case
archival pre-treatment biopsy tissue and post-chemotherapy tumor specimens were immunohistochemically
stained against CD31 and VEGF, as markers of angiogenic proliferation both in newly diagnosed primary
osteosarcoma and after multidrug chemotherapy including high-dose methotrexate (HDMTX). The correlation
between clinicopathological parameters and the degree of tumor VEGF and CD31 expression was statistically
assessed using the c2 test verified with Yates’ test for comparison of two groups. Significance was set at p < 0,05.
Results: Expression of VEGF was positive in 11 cases/16 of cases at diagnosis. Moreover, 8 cases/16 untreated
osteosarcomas were CD31-negative, but the other 8 showed an high expression of CD31. VEGF expression in
viable tumor cells after neoadjuvant chemotherapy was observed in all cases; in particular, there was an increased
VEGF expression (post-chemotherapy VEGF - biopsy VEGF) in 11 cases/16. CD31 expression increased in 11 cases/16
and decreased in 3 cases after chemotherapy. The data relating to the change in staining following chemotherapy
appear statistically significant for VEGF expression (p < 0,05), but not for CD31 (p > 0,05).
Conclusions: Even if the study included few patients, these results confirm that VEGF and CD31 expression is
affected by multidrug chemotherapy including HDMTX. The expression of angiogenic factors that increase
microvessel density (MVD) can contribute to the penetration of chemotherapeutic drugs into the tumor in the
adjuvant stage of treatment. So VEGF could have a paradoxical effect: it is associated with a poor outcome but it
could be a potential target for anti-angiogenic therapy.