Identification of vitamin D3target genes in human breast cancer tissue.
Multiple epidemiological studies have shown that high vitamin D3status is strongly associated with improved breast cancer survival. To determine the molecular pathways influenced by 1 alpha, 25-dihydroxyvitamin D3(1,25D) in breast epithelial cells we isolated RNA from normal human breast and cancer tissues treated with 1,25D in an ex vivo explant system. RNA-Seq revealed 523 genes that were differentially expressed in breast cancer tissues in response to 1,25D treatment, and 127 genes with altered expression in normal breast tissues. GoSeq KEGG pathway analysis revealed 1,25D down-regulated cellular metabolic pathways and enriched pathways involved with intercellular adhesion. The highly 1,25D up-regulated target genes CLMN, SERPINB1, EFTUD1, and KLK6were selected for further analysis and up-regulation by 1,25D was confirmed by qRT-PCR analysis in breast cancer cell lines and in a subset of human clinical samples from normal and cancer breast tissues. Ketoconazole potentiated 1,25D-mediated induction of CLMN, SERPINB1, and KLK6 mRNA through inhibition of 24-hydroxylase (CYP24A1) activity. Elevated expression levels of CLMN, SERPINB1, and KLK6 are associated with prolonged relapse-free survival for breast cancer patients. The major finding of the present study is that exposure of both normal and malignant breast tissue to 1,25D results in changes in cellular adhesion, metabolic pathways and tumor suppressor-like pathways, which support epidemiological data suggesting that adequate vitamin D3levels may improve breast cancer outcome.
School of Medicine, University of Adelaide, Adelaide, SA, Australia.
Lei
L
Sheng
School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, SA, Australia.
Paul H
PH
Anderson
School of Medicine, University of Adelaide, Adelaide, SA, Australia.
Andrew G
AG
Turner
School of Medicine, University of Adelaide, Adelaide, SA, Australia.
Kathleen I
KI
Pishas
SA Pathology, Adelaide, SA, Australia.
Deepak J
DJ
Dhatrak
School of Medicine, University of Adelaide, Adelaide, SA, Australia.
Peter G
PG
Gill
School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, SA, Australia; SA Pathology, Adelaide, SA, Australia.
Howard A
HA
Morris
School of Medicine, University of Adelaide, Adelaide, SA, Australia. Electronic address: david.callen@adelaide.edu.au.
David F
DF
Callen
D000970:Antineoplastic Agents / Q000494:pharmacology*; D001940:Breast / Q000378:metabolism; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000601:surgery; D002117:Calcitriol / Q000494:pharmacology; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002762:Cholecalciferol / Q000378:metabolism*; D018572:Disease-Free Survival; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007610:Kallikreins / Q000378:metabolism; D007654:Ketoconazole / Q000494:pharmacology; D061986:MCF-7 Cells; D008565:Membrane Proteins / Q000378:metabolism; D009364:Neoplasm Recurrence, Local; D010445:Peptide Elongation Factors / Q000378:metabolism; D017415:Ribonucleoprotein, U5 Small Nuclear / Q000378:metabolism; D017423:Sequence Analysis, RNA; D015843:Serpins / Q000378:metabolism; D015398:Signal Transduction; D014158:Transcription, Genetic; D015854:Up-Regulation; D065668:Vitamin D3 24-Hydroxylase / Q000037:antagonists & inhibitors; D000096985:Calponins
D000970:Antineoplastic Agents; D002117:Calcitriol; D002762:Cholecalciferol; D007610:Kallikreins; D007654:Ketoconazole; D008565:Membrane Proteins; D010445:Peptide Elongation Factors; D017415:Ribonucleoprotein, U5 Small Nuclear; D015843:Serpins; D065668:Vitamin D3 24-Hydroxylase; C435688:CLMN protein, human; D000096985:Calponins; C000599132:EFL1 protein, human; C075177:SERPINB1 protein, human; C585549:CYP24A1 protein, human; C103551:KLK6 protein, human
Breast cancer; Genomics; Mammary tissue; RNA-Seq; Vitamin D(3)