The anti-breast cancer drug tamoxifen alters Ca2+ movement in Chinese hamster ovary (CHO-K1) cells.
The anti-breast cancer drug tamoxifen has recently been shown to cause an increase in intracellular free-Ca(2+) concentrations ([Ca(2+)](i)) in renal tubular cells, breast cells and bladder cells. Because tamoxifen is known to alter ovary function in human patients and in rats, the present study was aimed at exploring whether tamoxifen could alter Ca(2+) movement in Chinese hamster ovary (CHO-K1) cells. Cytosolic free-Ca(2+) levels in populations of cells have been explored by using fura-2 as a fluorescent Ca(2+) indicator. Tamoxifen at concentrations above 1 micro M increased [Ca(2+)](i) in a concentration-dependent manner with an EC(50) value of 8 micro M. The Ca(2+) signal was reduced by removing extracellular Ca(2+), but was not affected by nifedipine, verapamil, diltiazem or ICI 182,780 (an estrogen receptor antagonist). Pretreatment with 1 micro M thapsigargin (an endoplasmic reticulum Ca(2+) pump inhibitor) to deplete the endoplasmic reticulum Ca(2+) abolished 10 micro M tamoxifen-induced Ca(2+) release. Neither inhibition of phospholipase C with 2 micro M U73122 nor depletion of ryanodine-sensitive Ca(2+) stores with 50 micro M ryanodine affected tamoxifen-induced Ca(2+) release. Cell proliferation assays using ELISA revealed that overnight incubation with 5-10 micro M tamoxifen inhibited cell proliferation by 20%, and 20 micro M tamoxifen killed all cells. Together, the results suggest that, in CHO-K1 cells, tamoxifen induced a [Ca(2+)](i) increase by causing store-Ca(2+) release from the endoplasmic reticulum in an phospholipase C-independent manner, and by inducing Ca(2+) influx. The action of tamoxifen appears to be dissociated from estrogen receptor activation. Longer incubation with tamoxifen (>5 micro M) was cytotoxic.
Jan
Chung-Ren
CR
Department of Medical Education and Research, Kaohsiung Veterans General Hospital, 813, Kaohsiung, Taiwan. crjan@isca.vghks.gov.tw
An-Jen
Chiang
C
Chang
Hong-Tai
HT
Roan
Cherng-Jau
CJ
Lu
Yih-Chau
YC
Jiann
Bang-Ping
BP
Ho
Chin-Man
CM
Huang
Jong-Khing
JK
D000818:Animals; D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology*; D016466:CHO Cells; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D020847:Estrogen Receptor Modulators / Q000494:pharmacology; D005456:Fluorescent Dyes; D016257:Fura-2; D013629:Tamoxifen / Q000494:pharmacology*; D013997:Time Factors; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors
D018931:Antineoplastic Agents, Hormonal; D002121:Calcium Channel Blockers; D020847:Estrogen Receptor Modulators; D005456:Fluorescent Dyes; D013629:Tamoxifen; D010738:Type C Phospholipases; D002118:Calcium; D016257:Fura-2