Fibroblast growth factor 2 (FGF‐2) and its receptors (FGFRs) are important regulators of bone cell function. Although FGF‐2 is a major modulator of bone cell function, its expression and regulation in human osteoblasts have not been investigated. We examined FGF‐2 messenger RNA (mRNA) expression and regulation in the human osteosarcoma MG‐63 cells. Northern analysis revealed that MG‐63 cells expressed FGF‐2 mRNA transcripts of 7, 4, 2.2, and 1.3 kilobases (kb). In the absence of serum, treatment with transforming growth factor beta (TGF‐β; 0.1‐10 ng/ml) increased all FGF‐2 mRNA transcripts. Maximal increase was seen with 1 ng/ml of TGF‐β. TGF‐β increased FGF‐2 mRNA expression within 2 h and this was sustained for 24 h. Phorbal myristate acetate (PMA; 1 μM) also increased FGF‐2 mRNA at 6 h. Time course studies showed that TGF‐β did not significantly alter FGFR1 or FGFR2 mRNA expression in MG‐63 cells. Western blotting with anti‐human FGF‐2 revealed that MG‐63 cells synthesize three isoforms of FGF‐2 protein of ∼18, 22/23, and 24 kDa, which were increased after either 6 h or 24 h of treatment with TGF‐β. Increased FGF‐2 mRNA and protein expression in response to TGF‐β was markedly reduced by the protein kinase A (PKA) inhibitor H‐89. Immunogold labeling of MG‐63 cells treated with TGF‐β showed increased labeling for FGF‐2 and FGFR2 in the nuclei. In contrast, TGF‐β treatment significantly decreased FGFR1 labeling in the nuclei. These data show that TGF‐β regulates FGF‐2 gene expression in human osteosarcoma cells. Furthermore, TGF‐β modulates the cellular localization of FGF‐2 and its receptors.