Abstract
Osteosarcoma (OS) is the most common primary pediatric malignancy of the bone having poor prognosis and long-term survival rates of less than 30% in patients with metastasis. MicroRNA-509 was reported to be downregulated in OS. We and others previously published that miR-509-3p can strongly attenuate cellular migration/invasion and sensitize ovarian cancer to cisplatin. Here, we show that overexpression of miR-509-3p inhibited migration of primary OS cell lines U2OS, HOS, and SaOS2 as well as metastatic derivatives 143B and LM7. miR-509-3p overexpression also inhibited proliferation and invasion of HOS and 143B cells and sensitized cells to cisplatin. Luciferase reporter assays using 3′-UTRs of predicted miR-509-3p targets associated with metastatic phenotypes revealed ARHGAP1 could be one of the downstream effectors of miR-509-3p in HOS. To find the global impact of miR-509-3p overexpression and cisplatin treatment we performed Reverse Phase Protein Analysis (RPPA). AXL, which has been reported to play a critical role in cisplatin resistance and confirmed as direct target of miR-509-3p was downregulated upon miR-509-3p treatment and further down-regulated upon miR-509-3p + cisplatin treatment. We propose that the miR-509-3p/AXL and miR-509-3p/ARHGAP1 axes have the potential to uncover new druggable targets for the treatment of drug resistant metastatic osteosarcoma.
Highlights
Osteosarcoma (OS) is the most common primary pediatric malignancy of the bone having poor prognosis and long-term survival rates of less than 30% in patients with metastasis
We examined the role of miR-509-3p in migration and invasion of OS cells in the presence or absence of the chemotherapeutic drug cisplatin and through microRNA target predictions and Reverse Phase Protein Array (RPPA) we identified downstream effectors of miR-509-3p
Zhang et al 2017, reported that miR-509-5p is downregulated in OS which suggests that miR-509-3p, which is expressed on the same hairpin as miR-509-5p is downregulated in OS. miR-509-3p has been established to inhibit proliferation and increase sensitivity to cisplatin in chemo-resistant ovarian cancer cells and influence the cell cycle, colony formation and migration of human epithelial lung and cervical cancer cell line[12,22,23]
Summary
Osteosarcoma (OS) is the most common primary pediatric malignancy of the bone having poor prognosis and long-term survival rates of less than 30% in patients with metastasis. To confirm actual binding of miR-509-3p to its predicted binding site on 3′-UTR of ARHGAP1 gene (Fig. 2E), we performed a luciferase reporter assay in HOS cells.
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