Linked Life Data

8253858

Source:http://linkedlifedata.com/resource/pubmed/id/8253858

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
1994-1-13
pubmed:abstractText
Lipid synthesis and secretion was measured in primary rat mammary epithelial cells cultured on basement matrix in medium supplemented with lactogenic hormones. The cells grew and differentiated to form alveolar-like structures reminiscent of lactating mammary gland. They synthesized abundant triacylglycerol, containing fatty acids characteristic of rat milk (C10:0-C14:0), using 14C-glucose, 14C-oleic acid or 14C-glycerol as precursors. Basal levels of triacylglycerol secretion were measured using 14C-oleic acid labeling; 1.3 +/- 0.3% of the labeled cellular triacylglycerol was secreted into the medium in 24 hours. Secreted lipid droplets were surrounded by a bilayer membrane with an electron-dense inner coat characteristic of fat globules secreted by the mammary gland. The rate of triglycerol secretion was increased to 998 +/- 98% of control (P < 0.01) by the addition of phorbol 12-myristate 13-acetate (PMA) in combination with staurosporine, a protein kinase inhibitor. Several other protein kinase inhibitors, when combined with PMA, also markedly stimulated secretion. Effective protein kinase inhibitors included sphingosine (has diverse cellular effects including the inhibition of protein kinase C; 13-fold increase in secretion), and KT5823 (a cGMP dependent protein kinase inhibitor; 5-fold increase). KT5720 (a cAMP-dependent protein kinase inhibitor) did not alter secretion. Kinase inhibitors were effective only in the presence of a phorbol ester. 4 alpha-phorbol-12,13-didecanoate, a phorbol ester which does not activate protein kinase C (PKC), could substitute for PMA. Lipid release was not mediated by disruption of cell-cell tight junctions, as EGTA did not release lipid. Based on these observations we suggest that two signals are needed to enable or stimulate lipid secretion in cultured rat mammary epithelial cells: 1) inhibition of a protein kinase and 2) a PKC-independent effect of phorbol ester. We have, for the first time, characterized a cell culture model suitable for studying lipid synthesis and secretion by mammary epithelial cells.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0021-9541
pubmed:author
pubmed:issnType
Print
pubmed:volume
157
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
469-80
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
1993
pubmed:articleTitle
Lipid synthesis and secretion by primary cultures of rat mammary epithelial cells.
pubmed:affiliation
Department of Nutrition, School of Public Health, University of North Carolina at Chapel Hill 27599-7400.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.