Effect of Carvacrol on Ca^(2+) Movement and Viability in PC3 Human Prostate Cancer Cells

Abstract

Carvacrol, a monoterpenic phenol compound, has been shown to possess various biological effects in different models. However, the effect of carvacrol on intracellular Ca²⁺ and its related physiology in human prostate cancer is unknown. This study explored the effect of carvacrol on cytosolic free Ca²⁺ levels ([Ca²⁺]i) and viability in PC3 human prostate cancer cells. Fura-2, a Ca²⁺- sensitive fluorescent dye, was used to assess [Ca²⁺]i. Cell viability was measured by the detecting reagent WST-1. Carvacrol at concentrations of 200-800 μM caused [Ca²⁺]i rises in a concentration-dependent manner. Removal of extracellular Ca²⁺ reduced carvacrol’s effect by approximately 60%. Carvacrol-induced Ca²⁺ entry was confirmed by Mn²⁺ entry-induced quench of fura-2 fluorescence, and was inhibited by approximately 30% by nifedipine, econazole, SKF96365, and the protein kinase C (PKC) inhibitor GF109203X. In Ca²⁺-free medium, treatment with the endoplasmic reticulum Ca²⁺ pump inhibitor thapsigargin (TG) abolished carvacrol-induced [Ca²⁺]i rises. Treatment with carvacrol also abolished TG-induced [Ca²⁺]i rises. Carvacrol-induced Ca²⁺ release from the endoplasmic reticulum was abolished by inhibition of phospholipase C (PLC). Carvacrol killed cells at concentrations of 200-600 μM in a concentration-dependent fashion. Chelating cytosolic Ca²⁺ with BAPTA/AM did not prevent carvacrol’s cytotoxicity. Together, in PC3 cells, carvacrol induced [Ca²⁺]i rises by inducing PLC-dependent Ca²⁺ release from the endoplasmic reticulum and Ca²⁺ entry via PKC-sensitive store-operated Ca²⁺ channels and other unknown channels. Carvacrol also induced Ca²⁺-dissociated cell death.

Exploring the impact of a naturally occurring sapogenin diosgenin on underlying mechanisms of Ca2+ movement and cytotoxicity in human prostate cancer cells

“…The Mn 2+ quenching data support that Ca 2+ influx occurred during diosgenin incubation. In PC3 cells, store‐operated Ca 2+ entry plays a critical role in Ca 2+ influx . Nifedipine, econazole, and SKF96365 are often applied to modulate store‐operated Ca 2+ entry in cellular studies .…”

“…As described previously, the measurement of cytotoxicity relied on the capacity of live cells to break down tetrazolium salts by dehydrogenases. Color density will develop proportionally to the amount of live cells.…”

“…In in vitro research of prostate cancer, the PC3 cell was often used because it grows rapidly in culture and produces [Ca 2+ ] i increases upon stimulation of different chemicals. In this cell, it has been reported that [Ca 2+ ] i rises and death can be evoked by chemicals including carvacrol, timolol, and thymol . To this end, our study applied fura‐2 as a Ca 2+ ‐sensitive probe to measure [Ca 2+ ] i in suspended cells in an effort to characterize the [Ca 2+ ] i rises, plot the concentration‐response relationship, and uncover the routes underlying diosgenin‐induced Ca 2+ release and Ca 2+ influx.…”

Ca2+ movement and cytotoxicity induced by the pyrethroid pesticide bifenthrin in human prostate cancer cells

“…[Ca 2þ ] i measurements [Ca 2þ ] i was measured as previously described. [18][19][20] Confluent cells grown on 6 cm dishes were trypsinized and made into a suspension in culture medium at a concentration of 10 6 cells per milliliter. Cell viability was determined by trypan blue exclusion.…”

“…The PC3 human prostate cancer cells were used because it produces measurable [Ca 2þ ] i rises upon pharmacological stimulation. It has been shown that in this cell, [Ca 2þ ] i rises and death can be evoked by stimulation with chemicals, such as timolol, 18 carvacrol, 19 and thymol. 20 To understand the physiological significance of this Ca 2þ signal, it is important to elucidate the mechanisms underlying the signal.…”