The effect of para (p)-cresol, an essential oil component, on calcium ion (Ca2+) signaling in human glioblastoma is unknown. The present study aimed to investigate how p-cresol influences intracellular Ca2+ levels ([Ca2+]i) and viability in DBTRG-05MG human glioblastoma cells. Cells were treated with p-cresol to assess its impact on cell viability and [Ca2+]i. Cell viability was evaluated using a WST-1 assay. [Ca2+]i was measured using a fluorescence-based Ca2+ indicator. Cells were loaded with the Ca2+-sensitive dye (fura-2), and fluorescence intensity was recorded before and after p-cresol treatment to determine changes in [Ca2+]i. p-Cresol induced concentration-dependent increases in [Ca2+]i between 50 and 150 µM. At 50-250 µM, p-cresol triggered cell death; this effect was reversed by pretreating the cells with the Ca2+ chelator BAPTA-AM. The removal of extracellular Ca2+ inhibited Ca2+ entry. p-Cresol-induced Ca2+ influx was confirmed by Mn2+-induced quenching of fura-2 fluorescence. Store-operated Ca2+ channel modulators SKF96365 and 2-aminoethoxydiphenyl borate and the protein kinase C inhibitor GF109203X inhibited p-cresol-induced Ca2+ entry, but voltage-gated Ca2+ channel blocker nifedipine did not. Treatment with the endoplasmic reticulum Ca2+ pump inhibitor thapsigargin in Ca2+-free medium inhibited p-cresol-induced [Ca2+]i rises; conversely, treatment with p-cresol decreased thapsigargin-induced [Ca2+]i rises. Furthermore, phospholipase C (PLC) inhibition with U73122 abolished p-cresol-induced [Ca2+]i rises. In DBTRG-05MG cells, p-cresol triggered Ca2+-associated cell death. The process involved the entry of Ca2+ through PKC-regulated store-operated Ca2+ channels and release of Ca2+ from the endoplasmic reticulum, which depends on PLC. Additionally, BAPTA-AM, which has Ca2+-chelating properties, may be a promising compound in preventing p-cresol-induced cytotoxicity, a potential breakthrough in neurotoxic research in glioblastoma cell model.
原文链接
分享
引用
