RCAN1.4 suppresses the osteosarcoma growth and metastasis via interfering with the calcineurin/NFAT signaling pathway

Calcipressin-1, also known as a regulator of calcineurin 1 (RCAN1), is one of the families of endogenous regulators of calcineurin activation and can specifically constrain the activity of calcineurin, but its function in osteosarcoma is still unknown. Firstly, we examined the protein level of RCAN1 in osteosarcoma specimens was lower than that of chondroma specimens. RCAN1.4 rather than RCAN1.1 had a higher endogenous protein level in six osteosarcoma cell lines by western blot. Further, we created stable RCAN1.4-deficient 143B and Hos cells using CRISPR-Cas9. RCAN1.4 loss promoted tumor growth in subcutaneous xenograft models. RCAN1.4 knockdown promoted tumor metastases to the lungs using intravenous metastasis models. Furthermore, we found that higher activity of calcineurin in RCAN1.4-deficient cells enhanced the nuclear translocation of NFATc1 to induce the cyclin D1 and MMPs expression. In addition, RCAN1.4 overexpression restrained osteosarcoma cell growth and invasion and inhibited the activity of calcineurin. Finally, we discovered that conditioned medium (20%) derived from RCAN1.4-deficient cells significantly promoted osteoclastogenesis, indicating Receptor Activator of Nuclear factor κB (RANK) signaling activation during osteosarcoma metastasis. In conclusion, RCAN1.4 may be a potential therapeutic target for osteosarcoma.