The protein-functionalized reduced graphene oxide (rGO) nanosheet is of great interest in stimuli-responsive drug delivery and controlled release applications. We developed doxorubicin (DOX)-loaded bovine serum albumin (BSA)-functionalized rGO (DOX-BSA-rGO) nanosheets. To investigate the reduction of BSA-functionalized GO nanosheets and drug loading efficiency, we used X-ray photoelectron spectroscopy (XPS) and UV-visible spectrophotometer analysis. DOX-BSA-rGO nanosheets exhibited dose-dependent cellular uptake without any cytotoxic effect. We also demonstrated near-infrared (NIR)-induced chemo-photothermal therapy of brain tumor cells treated with DOX-BSA-rGO nanosheets. Therefore, this DOX-BSA-rGO nanosheet could be a powerful tool for chemo-photothermal therapy applications.