Essential oil extract p‑cresol effect on Ca<sup>2+</sup> signaling and its underlying mechanism in DBTRG‑05MG human glioblastoma cells

Chou,Pin-Hao; Su,Chun-Lang; Fu,Shih-Hau; Schleip,Robert; Liang,Wei-Zhe

doi:10.3892/br.2025.1958

Essential oil extract p‑cresol effect on Ca²⁺ signaling and its underlying mechanism in DBTRG‑05MG human glioblastoma cells

Authors:
- Pin-Hao Chou
- Chun-Lang Su
- Shih-Hau Fu
- Robert Schleip
- Wei-Zhe Liang
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Affiliations: Department of Acupressure Technology, Chung Hwa University of Medical Technology, Tainan 717302, Taiwan, R.O.C., Chung Jen Junior College of Nursing, Health Science and Management, Chiayi 60077, Taiwan, R.O.C., European Rolfing Association, Medical Faculty of Ulm University, D‑89081 Ulm, Germany, Department of Pharmacy and Master Program, College of Pharmacy and Health Care, Tajen University, Yanpu, Pingtung 907101, Taiwan, R.O.C.
Published online on: March 4, 2025 https://doi.org/10.3892/br.2025.1958
Article Number: 80

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Abstract

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.

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