Alterations in metal toxicity and metal-induced metallothionein gene expression elicited by growth medium calcium concentration.
The calcium content of the growth medium has been shown to influence the growth and differentiation of primary epithelial cells in culture. The goal of the present study was to determine if growth medium calcium concentration could influence the susceptibility to metal toxicity and metallothionein gene expression of an immortalized human prostate-derived epithelial cell line (RWPE-1). The RWPE-1 cell line was grown in medium containing either 0.1 or 1.4 mM calcium. Confluent cells were exposed to either Zn(+2) (50, 100, or 150 microM) or Cd(+2) (3, 6, or 12 microM) for 13 days, and cell toxicity and MT gene expression were determined along the time course of exposure. It was demonstrated that the calcium content of the growth medium had a marked influence on Zn(+2) toxicity and a lesser but significant effect on Cd(+2) toxicity to the RWPE-1 cells. Calcium concentration of the growth medium was also shown to alter the accumulation of MT-1/2 protein and MT-1E, MT-1X, and MT-2A mRNAs. It was shown that MT-1/2 protein was markedly increased for metal-exposed cells grown in medium containing 0.1 mM calcium; however, the increased expression did not cause an increase in the resistance of the cells to Zn(+2) or Cd(+2) exposure. These observations show that growth medium calcium concentration can influence metal toxicity and the pattern of expression of the MT mRNAs and protein for RWPE-1 cells. The results suggest that caution should be exercised when comparing toxicological responses between cell lines that may be grown in growth formulations differing in calcium concentration.
Singh
Rajendra K
RK
Department of Pathology, School of Medicine and Health Sciences, University of North Dakota, 501 N. Columbia Road Stop 9037, Grand Forks, ND 58202, USA.
Albrecht
Amy L
AL
Somji
Seema
S
Sens
Mary Ann
MA
Sens
Donald A
DA
Garrett
Scott H
SH
D001483:Base Sequence; D002104:Cadmium / Q000633:toxicity*; D002118:Calcium / Q000378:metabolism*; D002461:Cell Line, Transformed; D003470:Culture Media; D017931:DNA Primers; D005786:Gene Expression Regulation / Q000187:drug effects*; D008668:Metallothionein / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D015032:Zinc / Q000633:toxicity*
D003470:Culture Media; D017931:DNA Primers; D012333:RNA, Messenger; D002104:Cadmium; D008668:Metallothionein; D015032:Zinc; D002118:Calcium