The carcinogen safrole increases intracellular free Ca 2+ levels and causes death in MDCK cells

Wei Chuan Chen, He Hsiung Cheng, Chun Jen Huang, Yih Chau Lu, I. Shu Chen, Shiuh Inn Liu, Shu Shong Hsu, Hong Tai Chang, Jong Khing Huang, Jin Shyr Chen, Chung Ren Jan*

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

11 Scopus citations


The effect of the carcinogen safrole on intracellular Ca 2+ movement in renal tubular cells has not been explored previously. The present study examined whether safrole could alter Ca 2+ handling in Madin-Darby canine kidney (MDCK) cells. Cytosolic free Ca 2+ levels ([Ca 2+ ] i ) in populations of cells were measured using fura-2 as a fluorescent Ca 2+ probe. Safrole at concentrations above 33 μM increased [Ca 2+ ] i in a concentration-dependent manner with an EC 50 value of 400 μM. The Ca 2+ signal was reduced by 90% by removing extracellular Ca 2+ , but was not affected by nifedipine, verapamil, or diltiazem. Addition of Ca 2+ after safrole had depleted intracellular Ca 2+ -induced dramatic Ca 2+ influx, suggesting that safrole caused store-operated Ca 2+ entry. In Ca 2+ -free medium, after pretreatment with 650 μM safrole, 1 μM thapsigargin (an endoplasmic reticulum Ca 2+ pump inhibitor) failed to release more Ca 2+ . Inhibition of phospholipase C with 2 μM U73122 did not affect safrole-induced Ca 2+ release. Trypan blue exclusion assays revealed that incubation with 650 μM safrole for 30 min did not kill cells, but killed 70% of cells after incubation for 60 min. Collectively, the data suggest that in MDCK cells, safrole induced a [Ca 2+ ] i increase by causing Ca 2+ release from the endoplasmic reticulum in a phospholipase C-independent fashion, and by inducing Ca 2+ influx via store-operated Ca 2+ entry. Furthermore, safrole can cause acute toxicity to MDCK cells.

Original languageEnglish
Pages (from-to)34-40
Number of pages7
JournalChinese Journal of Physiology
Issue number1
StatePublished - 2007
Externally publishedYes


  • Ca
  • Ca stores
  • Fura-2
  • MDCK
  • Renal tubular cells
  • Safrole


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