Rationale: Extracellular vesicles (EVs) have emerged as novel mediators of cell-to-cell communication that are capable of the stable transfer of therapeutic
microRNAs (
miRNAs), and thus, EVs hold immense promise as a
miRNA delivery system for
cancer therapy. Additionally, as
miRNA-containing EVs are secreted into circulation,
miRNAs contained within plasma EVs may represent ideal
biomarkers for diseases. The objective of this study was to characterize a potential
tumor suppressor
miRNA, miR-101, and explore the potential of miR-101 delivery via EVs for in vivo
therapy of metastatic
osteosarcoma as well as the potential value of plasma EV-packaged miR-101 (EV-miR-101) level for predicting
osteosarcoma metastasis. Methods: The relationship of miR-101 expression and
osteosarcoma progression was investigated in
osteosarcoma specimens by in situ hybridization (ISH), and the potential inhibitory effect of miR-101 was further investigated using in vivo models. Using prediction software analysis, the mechanism of action of miR-101 in
osteosarcoma was explored using quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blotting and dual-
luciferase assay. Adipose tissue-derived mesenchymal stromal cells (AD-MSCs) were transduced with lentiviral particles to obtain miR-101-enriched EVs. A Transwell assay and lung
metastasis models of
osteosarcoma were used to observe the effect of miR-101-enriched EVs on
osteosarcoma invasiveness and
metastasis. Detection of plasma EV-miR-101 levels was carried out in
osteosarcoma patients and healthy controls by qRT-PCR. Results: miR-101 expression was markedly lower in metastatic
osteosarcoma specimens compared to non-metastatic specimens. Significantly fewer metastatic lung nodules were formed by Saos-2 cells overexpressing miR-101 and SOSP-9607 cells overexpressing miR-101 injected into mice. With increased miR-101 expression,
B cell lymphoma 6 (BCL6)
mRNA and
protein expression levels were reduced, and miR-101 was found to exert its effects by directly targeting BCL6. AD-MSCs were successfully engineered to secrete miR-101-enriched EVs. Once taken up by
osteosarcoma cells, these EVs showed suppressive effects on cell invasion and migration in vitro, and systemic administration of these EVs effectively suppressed
metastasis in vivo with no significant side effects. Finally, the EV-miR-101 level was lower in
osteosarcoma patients than in healthy controls and even lower in
osteosarcoma patients with
metastasis than in those without
metastasis. Conclusion: Our data support the function of miR-101 as a
tumor suppressor in
osteosarcoma via downregulation of BCL6. AD-MSC derived miR-101-enriched EVs represent a potential
innovative therapy for metastatic
osteosarcoma. EV-miR-101 also represents a promising circulating
biomarker of
osteosarcoma metastasis.