Role of the cytoskeleton in laminin induced mammary gene expression.
The differentiation of rat mammary epithelial cells is characterized both by morphologic changes and by the expression of a group of milk protein genes. We have previously shown that by culturing these cells on the basement membrane glycoprotein laminin, the synthesis of the milk proteins, transferrin, alpha-casein, and alpha-lactalbumin is induced. In order to determine if this effect is mediated through the cytoskeleton, we have treated these cells with cytochalasin D and colchicine. Treatment with cytochalasin D or colchicine for 24 h inhibits the accumulation of alpha-casein, transferrin, and alpha-lactalbumin without significant effect on general protein synthesis. Pulse chase studies show that cytochalasin D does not alter the intracellular turnover of alpha-casein or transferrin. Additionally, treatment with cytochalasin D causes an early (within 1 h) increase in secretion of alpha-casein and transferrin suggesting that the actin cytoskeleton provides a meshwork for secretory vesicles. The disruption of this network enhances the secretion of preformed proteins. However, long term (24 h) treatment with cytochalasin D inhibits synthesis of these milk proteins. Northern blot analysis indicates that treatment with cytochalasin D or colchicine inhibits the laminin induced increase in alpha-casein, alpha-lactalbumin, and transferrin mRNAs. These studies indicate that the major effect of the cytoskeleton on laminin induced milk protein gene expression occurs at the level of accumulation of mRNAs for these proteins. We conclude that the expression of laminin induced milk protein gene expression in primary rat mammary cultures depends on the integrity of the actin and microtubule cytoskeleton.
Blum
J L
JL
Department of Internal Medicine, Simpson Memorial Research Institute, University of Michigan, Ann Arbor 48109.
Wicha
M S
MS
D000818:Animals; D002364:Caseins / Q000235:genetics; D002478:Cells, Cultured; D003078:Colchicine / Q000494:pharmacology; D015638:Cytochalasin D; D003572:Cytochalasins / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D004848:Epithelium / Q000378:metabolism / Q000648:ultrastructure; D005260:Female; D005786:Gene Expression Regulation; D005796:Genes / Q000187:drug effects*; D007768:Lactalbumin / Q000235:genetics; D007797:Laminin / Q000494:pharmacology*; D008321:Mammary Glands, Animal / Q000378:metabolism*; D008894:Milk Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011919:Rats, Inbred Strains; D014158:Transcription, Genetic / Q000187:drug effects*; D014168:Transferrin / Q000235:genetics
D002364:Caseins; D003572:Cytochalasins; D007797:Laminin; D008894:Milk Proteins; D012333:RNA, Messenger; D014168:Transferrin; D015638:Cytochalasin D; D007768:Lactalbumin; D003078:Colchicine