Hax-1 Regulates Radiation-Induced Mitochondrial-Dependent Apoptosis of Uveal Melanoma Cells through PI3K/AKT/eNOS Pathway

Abstract

Uveal melanoma is an aggressive skin cancer that remains insurmountable and is accompanied by inferior prognostic results. The proliferative and survival mechanisms of uveal melanoma cells need to be further investigated to improve the treatment of uveal melanoma. According to reports, HAX-1 is an antiapoptotic protein vital for multiple malignancies. Nevertheless, the role and causal link of HAX-1 in uveal melanoma are still elusive. The survival diversity of uveal melanoma sufferers with diverse haX-1 expressing levels was studied by TCGA database. Patients in the riskhigh group exhibited greater levels of HAX-1 in contrast to the risklow group, and individuals with higher HAX-1 levels displayed inferior survival times. The outcomes of CCK-8 and clonogenesis revealed that the proliferative rate of haX-1 knockout cells was slower. The result of scratch experiment shows that the ability of scratch recovery after HAX-1 is reduced. Transwell migration and tumor cell pelletization experiments showed that siHAX-1 significantly reduced cell migration and tumor cell pelletization. After haX-1 was knocked out, the loss of MMP was decreased, the transfer of CyT C was elevated, and the protein expression of Bax, Caspase 3, and Bcl2 was elevated, suggesting that mitochondria-induced apoptosis was increased. Sihax-1 treatment remarkably decreased the phosphonation of phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR)/endothelial NO synthase (eNOS) in mum-2B and C918. Pretreatment with LY294002 significantly restored iHAX-1-induced decline in PI3K/AKT/mTOR/eNOS phosphorylation. Therefore, our results suggest that haX-1 induces radiation-dependent apoptosis of UM cells via the PI3K/AKT/eNOS signal path.