Osteosarcoma is a malignant bone tumor, and clinically detectable metastases can be detected in ~ 15–20% of patients when they seek medical advice; patients with metastatic disease have extremely poor prognosis. Here, we examined the involvement of the microRNA miR‐505 in osteosarcoma. Eighty‐four patients seeking treatment for osteosarcoma were included in the study group (SG), and 63 healthy subjects were allocated to the control group (CG). Normal human bone cells MG‐63 and U20S cells were transfected with miR‐505 mimics, miR‐NC, HMGB1 RNA for targeted inhibition (si‐HMGB1), and si‐NC to examine the effects on HMGB1 expression. Cell proliferation, invasion, and apoptosis were measured using CCK‐8, scratch assays, and flow cytometry (FCM), respectively, and the relationship between miR‐505 and HMGB1 was determined using the dual‐luciferase reporter assay. In patient tissues and serum, miR‐505 was expressed at a low level, and HMGB1 was expressed at a high level, with an area under curve of > 0.9. Furthermore, the expression of miR‐505 and HMGB1 in tissues had a positive association with that in the serum, whereas the expression of miR‐505 had a negative association with that of HMGB1 in tissues only. Overexpression of miR‐505 and silencing of HMGB1 suppressed the proliferation, migration, and invasion of osteosarcoma cells and increased the rate of apoptosis, whereas the co‐transfected miR‐505 mimics + si‐HMGB1 demonstrated a more significant inhibitory effect on the proliferation and invasion of osteosarcoma cells and a higher apoptosis rate. miR‐505 may inhibit the proliferation and invasion and promote apoptosis of osteosarcoma cells by targeting and suppressing HMGB1.