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Department of Experimental Pathology, Roswell Park Cancer Institute, Buffalo, NY 14263;
*
Dipartimento di Biologia Sperimentale, Universita degli Studi di Cagliari, Cittadella Universitaria, 09042 Monserrato, Cagliari, Italy;
Division of Laboratory Research, AMC Cancer Research Center, Denver, CO 80214;
**
Department of Food Science and
Department of Animal Science, Cornell University, Ithaca, NY 14853
2To whom correspondence should be addressed.
Conjugated linoleic acid (CLA) is a potent cancer preventive agent in animal models. To date, all of the in vivo work with CLA has been done
with a commercial free fatty acid preparation containing a mixture of
c9,t11-, t10,c12- and c11,t13-isomers, although CLA in food is
predominantly (80–90%) the c9,t11-isomer present in triacylglycerols. The objective of this study was to determine whether
a high CLA butter fat has biological activities similar to those of the mixture of free fatty acid CLA isomers. The following four different
endpoints were evaluated in rat mammary gland: 1) digitized image analysis of epithelial mass in mammary whole mount;
2) terminal end bud (TEB) density; 3) proliferative activity of TEB cells as determined by proliferating cell
nuclear antigen immunohistochemistry; and 4) mammary cancer prevention bioassay in the methylnitrosourea model. It should be
noted that TEB cells are the target cells for mammary chemical carcinogenesis. Feeding butter fat CLA to rats during the time of
pubescent mammary gland development reduced mammary epithelial mass by 22%, decreased the size of the TEB population by 30%, suppressed the
proliferation of TEB cells by 30% and inhibited mammary tumor yield by 53% (P < 0.05). Furthermore, all of the above
variables responded with the same magnitude of change to both butter fat CLA and the mixture of CLA isomers at the level of CLA (0.8%)
present in the diet. Interestingly, there appeared to be some selectivity in the uptake or incorporation of c9,t11-CLA over
t10,c12-CLA in the tissues of rats given the mixture of CLA isomers. Rats consuming the CLA-enriched butter
fat also consistently accumulated more total CLA in the mammary gland and other tissues (four- to sixfold increases) compared with those
consuming free fatty acid CLA (threefold increases) at the same dietary level of intake. We hypothesize that the availability of vaccenic acid
(t11–18:1) in butter fat may serve as the precursor for the endogenous synthesis of CLA via the 
9-desaturase reaction.
Further studies will be conducted to investigate other attributes of this novel dairy product.
KEY WORDS: • conjugated linoleic acid • butter fat • mammary gland morphogenesis • mammary cancer prevention • tissue CLA isomers • rats
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