Background:Osteosarcoma (OS) is the primary malignant bone tumor that most commonly affects children, adolescents, and young adults. MicroRNA-34a (miR-34a) is involved in tumor metastasis and may be a prognostic marker for patients with cancer. The aim of the present study was to explore the role of miR-34a in patients with OS. The underlying associations between miR-34a expressions and metastasis, recurrence as well as and prognosis were comprehensively analyzed in OS patients.Methods:Reverse transcriptase quantitative PCR (RT-qPCR) was used to investigate serum level of miR-34a between clinical OS patients (n = 162) and age-matched healthy controls (n = 162). Expression of miR-34a in OS tissues and adjacent tissues was analyzed using RT-qPCR. RT-qPCR was used to compare the serum level of miR-34a in patients with OS before and after chemotherapy. Multivariate Cox-regression analysis was used to analyze the association between serum level of miR-34a and chemotherapy resistance, overall survival, as well as recurrence and prognosis of OS patients. Five-year recurrence and survival were estimated using Kaplan–Meier curves.Results:Serum level of miR-34a was downregulated in OS patients (n = 86) compared to age-matched healthy controls (n = 86). Expression of miR-34a was downregulated in OS tissue compared to adjacent tissues in clinical patients. The expression of serum miR-34a before and after chemotherapy was positively correlated with the expression of miR-34a in the corresponding tissues. Expression of miR-34a was higher in the group where chemotherapy was effective than that patient where chemotherapy was ineffective. Expression of miR-34a was negatively associated with chemotherapy resistance of OS patients. High serum levels of miR-34a were associated with longer overall survival in OS patients and lower metastasis. Multivariate Cox-regression analysis identified miR-34a serum level with potential prognostic significance.Conclusion:The expression level of serum miR-34a in patients with OS is closely related to the chemotherapy resistance, metastasis, recurrence, and survival of osteosarcoma, which can be used as one of the potential biomarkers and prognosis for the treatment of OS patients. Therefore, miR-34a may be a potential molecular for prediction of the efficacy of chemotherapy and prognosis in OS patients.