Neuroblastoma (NB) is a childhood cancer with a low survival rate and great metastatic potential. Calreticulin (CRT), an endoplasmic reticulum chaperone protein, has been identified as one of the prognosis factor for NB patients. Our previous study demonstrated that CRT promotes neuronal differentiation through the upregulation of VEGF-A in NB cell lines. Inhibition of VEGFR-1 signaling by blocking antibodies significantly suppressed the expressions of neuronal markers. These results suggested that VEGF-VEGFR signaling may play important roles in the differentiation of NB cells. In this study, we further confirmed our hypothesis through xenograft model and clinical analysis from NB patients. In xenograft experiments, CRT inducible stNB-V1 cells were injected subcutaneously. Doxcycline treated in the drinking water effectively inhibited the tumor growth. In addition, the mRNA and protein levels of VEGF-A and differentiation marker GAP-43 were upregulated by induced CRT expression. However, no significant correlation between the expression levels of VEGF-A and CD31, a marker of endothelial cells, was observed which suggested a novel mechanism of VEGF-A participating in NB tumorigenesis through an angiogenesis-independent pathway. From NB patients samples, the mRNA expression levels of CRT and VEGF-A were positive correlated. Furthermore, positive VEGF-A expression by immunostaining of NB tumors was found to correlate well with histological grade of differentiation and predicted a favorable prognosis. In conclusion, our findings suggested that CRT may promote neuronal differentiation of neuroblastoma through regulating VEGF-A expression. However, the underlying mechanisms still need further investigation.