title: RANKL expression in normal and malignant breast tissue responds to progesterone and is up-regulated during the luteal phase.
abstract: The receptor activator of nuclear factor-κB ligand (RANKL) acts as a paracrine factor in progesterone-induced mammary epithelial proliferation and tumorigenesis. This evidence comes mainly from mouse models. Our aim was to examine whether RANKL expression in human normal and malignant breast is under the control of progesterone throughout the menstrual cycle. Breast epithelial samples were obtained by random fine needle aspiration (rFNA) of the contralateral unaffected breasts (CUB) of 18 breast cancer patients, with simultaneous serum hormone measurements. Genes correlated with serum progesterone levels were identified through Illumina microarray analysis. Validation was performed using qRT-PCR in rFNA samples from CUB of an additional 53 women and using immunohistochemistry in tissue microarrays of 61 breast cancer samples. Expression of RANKL, DIO2, and MYBPC1 was correlated with serum progesterone in CUB, and was significantly higher in luteal phase. RANKL and MYBPC1 mRNA expression were highly correlated between CUB and matched tumor samples. RANKL protein expression was also significantly increased in the luteal phase and highly correlated with serum progesterone levels in cancer samples, especially in hormone receptor positive tumors. The regulatory effects of progesterone on the expression of RANKL, DIO2, and MYBPC1 were confirmed in three-dimensional cultures of normal breast organoids. In normal breast and in breast cancer, RANKL mRNA and protein expression fluctuate with serum progesterone with highest levels in the luteal phase, suggesting that RANKL is a modulator of progesterone signaling in normal and malignant breast tissue and a potential biomarker of progesterone action and blockade.
authors.affiliation: Department of Surgery, Northwestern University Feinberg School of Medicine, 303 E. Superior Street Lurie 4-111, Chicago, IL, 60611, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Hu
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.forename: Akash
authors.initials: A
authors.lastname: Gupta
authors.forename: Ali
authors.initials: A
authors.lastname: Shidfar
authors.forename: Daniel
authors.initials: D
authors.lastname: Branstetter
authors.forename: Oukseub
authors.initials: O
authors.lastname: Lee
authors.forename: David
authors.initials: D
authors.lastname: Ivancic
authors.forename: Megan
authors.initials: M
authors.lastname: Sullivan
authors.forename: Robert T
authors.initials: RT
authors.lastname: Chatterton
authors.forename: William C
authors.initials: WC
authors.lastname: Dougall
authors.forename: Seema A
authors.initials: SA
authors.lastname: Khan
mesh_terms: D000328:Adult; D000368:Aged; D044963:Biopsy, Fine-Needle; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology; D063646:Carcinogenesis; D002352:Carrier Proteins / Q000096:biosynthesis; D004958:Estradiol / Q000097:blood; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007453:Iodide Peroxidase / Q000096:biosynthesis; D008183:Luteal Phase / Q000235:genetics; D042361:Mammary Glands, Human / Q000378:metabolism / Q000473:pathology; D008597:Menstrual Cycle / Q000378:metabolism; D008875:Middle Aged; D061251:Primary Cell Culture; D011374:Progesterone / Q000097:blood*; D053245:RANK Ligand / Q000096:biosynthesis* / Q000097:blood / Q000235:genetics; D000097564:Iodothyronine Deiodinase Type II; D000099163:Myosin Binding Protein C
chemical_list: D002352:Carrier Proteins; D004958:Estradiol; D007453:Iodide Peroxidase; D011374:Progesterone; D053245:RANK Ligand; D000097564:Iodothyronine Deiodinase Type II; D000099163:Myosin Binding Protein C; C505688:TNFSF11 protein, human