JS-K—a NO prodrug—inhibits migration, invasion, and bone metastasis of renal cell carcinoma through the NF-κB signaling pathway

Background O2-(2, 4-dinitrophenyl) 1-[(4-ethoxycarbonyl) piperazin-1-yl] diazen-1-ium-1, 2-diolate (JS-K)—a nitric oxide prodrug—inhibits the proliferation and migration of breast cancer, non-small cell lung cancer, and liver cancer cells. However, its mechanism of action in renal cell carcinoma (RCC) remains elusive. This study seeks to investigate the effect of JS-K on RCC migration, invasion, and bone metastasisand elucidate the underlying molecular mechanisms. Methods Human RCC cell lines were treated with different concentrations of JS-K, the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) inhibitor Bay-11-7082, or receptor activator of nuclear factor-kappa beta ligand (RANKL), per experimental requirements. In vitro migration, invasion, adhesion, and bone metastasis were assessed by the wound healing assay, Transwell assay, and hanging drop assay. Western blotting and immunofluorescence staining were conducted to evaluate the expression of specific proteins. Changes in gene expression were analyzed by RNA sequencing and quantitative polymerase chain reaction. Results JS-K treatment inhibited the epithelial-mesenchymal transition and extracellular matrix remodeling of RCC in a concentration-dependent manner. Subsequently, the migration, invasion, bone metastasis, and adhesion abilities of RCC cells significantly decreased after JK-S treatment. At the molecular basis, JS-K upregulated nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha while inhibiting the nuclear translocation of p65 and p50, suggesting its involvement in the NF-κB signaling pathway. Pharmacological inhibition of NF-κB with either Bay-11-7082 or RANKL influenced the effects of JS-K on RCC migration, invasion, and bone metastasis. Conclusion JS-K treatment inhibits RCC migration, invasion, and bone metastasis by targeting the NF-κB signaling pathway.