Neuroglobin (NGB) is a characterized heme globin that is widely expressed in the vertebrate central and peripheral nervous system as well as retina and endocrine tissues. However, to date, it is not determined whether functional NGB is expressed in cells of glia origin. In this study, we aimed to explore the detailed expression of NGB in a rat astrocytoma cell line (C6) and human astrocytoma cell line (U251) by reverse transcription-polymerase chain reaction, immunofluorescence, and Western blotting, and to detect the expression of NGB in human astrocytoma tissues by an immunohistochemical method. We found that NGB was present in a rat astrocytoma cell line (C6), human astrocytoma cell line (U251), and human astrocytoma tissues. The expression and potential roles of NGB in astrocytomas may provide insight into the mechanisms of tumor cells to adapt and survive in hypoxic microenvironments and also represent a novel therapeutic approach to astrocytomas.