Cellular retinol-binding protein I, a regulator of breast epithelial retinoic acid receptor activity, cell differentiation, and tumorigenicity.
BACKGROUND
Retinoic acid receptor (RAR) activation induces cell differentiation and may antagonize cancer progression. Cellular retinol-binding protein I (CRBP-I) functions in retinol storage and its expression is lower in human cancers than in normal cells. We hypothesized that retinol storage might be linked to RAR activation and thus that lowered CRBP-I function might impair RAR activity and cell differentiation.


METHODS
Sarcoma virus 40-immortalized human mammary epithelial cells (MTSV1-7) devoid of CRBP-I were transfected with wild-type CRBP-I or CRBP-I point mutants with low RA binding affinity. The subcellular localization of CRBP-I was investigated in these cells and in wild-type or CRBP-I null mouse mammary epithelial cells (MECs), using indirect immunofluorescence and sucrose gradient fractionation. RAR activity was assessed using reporter gene assays. Acinar differentiation and in vivo tumor growth were assessed in reconstituted basement membrane and athymic mice, respectively.


RESULTS
In cells expressing wild-type CRBP-I but not the CRBP-I mutants, CRBP-I was found mainly in lipid droplets, the retinol storage organelle, and this localization was associated with promotion of retinol storage by wild-type CRBP-I only. RAR activity was higher and acinar differentiation was observed in cells expressing wild-type but not mutant CRBP-I. RAR antagonist treatment blocked and chronic RA treatment mimicked, the CRBP-I induction of cell differentiation. Finally, CRBP-I suppressed tumorigenicity in athymic mice.


CONCLUSIONS
Physiologic RAR activation is dependent on CRBP-I-mediated retinol storage, and CRBP-I downregulation chronically compromises RAR activity, leading to loss of cell differentiation and tumor progression.
Farias
Eduardo F
EF
Department of Medicine, Mount Sinai School of Medicine, Annenberg Bldg., Rm 24-74, One Gustave L. Levy Place, New York, NY 10029, USA. eduardo.farias@mssm.edu
Ong
David E
DE
Ghyselinck
Norbert B
NB
Nakajo
Shigeo
S
Kuppumbatti
Yuvarani S
YS
Mira y Lopez
Rafael
R
D000818:Animals; D001943:Breast Neoplasms / Q000378:metabolism*; D002454:Cell Differentiation; D018450:Disease Progression; D015536:Down-Regulation; D004847:Epithelial Cells / Q000378:metabolism*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D015972:Gene Expression Regulation, Neoplastic; D017930:Genes, Reporter; D006801:Humans; D042361:Mammary Glands, Human / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008819:Mice, Nude; D017354:Point Mutation; D018168:Receptors, Retinoic Acid / Q000235:genetics / Q000378:metabolism*; D012177:Retinol-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D054840:Retinol-Binding Proteins, Cellular; D013395:Sucrose / Q000378:metabolism; D014162:Transfection
C518456:RBP1 protein, human; D018168:Receptors, Retinoic Acid; D012177:Retinol-Binding Proteins; D054840:Retinol-Binding Proteins, Cellular; D013395:Sucrose