Genetic alternations, such as mutations caused inactivities of tumor suppressor gene, have been identified in a wide variety of tumors, including osteosarcoma. Osteosarcoma is the most frequent primary malignant bone tumor that occurs in the extremities of young adolescents in most cases. Because of the high frequent occurrence of this type of tumor in hereditary retinoblastoma patients, involvement of the Rbl gene mutations was suspected in the development of osteosarcoma, and a few reports have shown alternations of the Rbl gene in osteosarcoma. We studied Rbl gene mutations in 9 osteosarcoma samples and one cell line (OS 732) to explore the types and mechanism of Rbl gene mutations in osteosarcoma.