Zinc-induced metallothionein synthesis by Caco-2 cells.
Caco-2 cells possess many morphological and biochemical characteristics of intestinal absorptive cells, including the ability to transport zinc. In the present study, metallothionein (MT) synthesis in response to increased levels of zinc was examined. Increased incorporation of [35S]cysteine into MTs was observed when excess ZnCl2 was added to the medium. The rate of MT synthesis was found to be concentration dependent. Also, induction of MT synthesis was greater early in the culture, before the cells were fully differentiated. Incubation of the monolayers with 65Zn and 200 microM zinc revealed that approximately 50% of the zinc incorporated into the cells was associated with MTs. The remainder was associated with large proteins as well as amino acids and small peptides. Actinomycin D and cycloheximide both inhibited the induction of MT synthesis, suggesting that the newly synthesized MTs are a result of expression of MT genes. Hence, Caco-2 cells, a model of intestinal absorptive cells, may be used to examine the role of MTs in zinc absorption.
Raffaniello
R D
RD
Department of Pediatrics, North Shore University Hospital-Cornell University Medical College, Manhasset, New York 11030.
Wapnir
R A
RA
D000818:Animals; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D003545:Cysteine / Q000378:metabolism; D003609:Dactinomycin / Q000494:pharmacology; D007408:Intestinal Absorption; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000187:drug effects; D008668:Metallothionein / Q000096:biosynthesis*; D015032:Zinc / Q000378:metabolism / Q000494:pharmacology*
D003609:Dactinomycin; D008668:Metallothionein; D003513:Cycloheximide; D015032:Zinc; D003545:Cysteine