The Small Non-coding RNA miR-16-1-3p Hampers Cancer Stem Cell Self-renewal and Invasiveness, Boosting Chemosensitivity by Adjusting TGF-β1 Signaling via MDM2/p53 Axis in Human Osteosarcoma.

Abstract

The TGF-{beta} signaling pathway has both tumor-suppressing and metastasis-promoting effects in cancer. However, the molecular determinants governing this switch remain unclear. Here, we explored the miR-16-1-3p/MDM2/p53 axis as a critical conductor of the TGF-{beta}-Smad pathway in osteosarcoma. Although miR-16-1-3p overexpression by itself markedly reduces proliferative and clonogenic potential of U2OS cells, when paired with TGF-{beta} treatment, it significantly increases arrest cells in G1 phase and nearly extinguishing the growth capability of these cells. MiR-16-1-3p overexpression inhibited TGF-induced actin remodeling and EMT featuring, significantly decreasing vimentin levels. TGF-{beta} enhances both 2D and 3D migration, but miR-16-1-3p overexpression, alone or with TGF-{beta}, strongly counteracts its pro-migratory effects. MiR-16-1-3p restored p53 stability by targeting MDM2, redirecting TGF-{beta}-Smad signaling toward p21 activation and proliferation inhibition while attenuating its EMT-promoting capacity. Administration of TGF-{beta} together with miR-16-1-3p dramatically increases the sensitivity of wild-type U2OS cells to cisplatin, exceeding that of TGF-{beta} therapy alone by more than an order of magnitude. Administering TGF-{beta} and miR-16-1-3p together significantly reduces the tumor nodule volume and Ki67 expression, while effectively eradicates metastases in the chicken chorioallantoic membrane (CAM) in vivo model. For the first time, our research demonstrates that miR-16-1-3p shifts TGF-{beta}1 signaling from a facilitator of metastasis to a promoter of anti-growth effects through MDM2 inhibition and p53 stabilization, effectively reducing the self-renewal and invasiveness of cancer stem cells in human osteosarcoma model. This process preserves TGF-{beta}s tumor-suppressive role while limiting its associated cancer risks.