Research paperProgestin-mediated activation of MAPK and AKT in nuclear progesterone receptor negative breast epithelial cells: The role of membrane progesterone receptors
Section snippets
1. Introduction
Progesterone (P4) is a steroid produced predominantly by the corpus luteum during estrous cycles and pregnancy, as well as the placenta during gestation. P4 elicits a number of different biological responses in females principally associated with establishment and maintenance of pregnancy, influencing several target tissues. In breasts, P4 promotes formation of lobular-alveolar structures during pregnancy, regulating normal breast physiology (Topper and Freeman, 1980). Specifically, P4
2.1. Cell culture
MCF12A and MCF10A human epithelial breast cells were purchased from ATCC. Cells were maintained in a humidified incubator at 37 °C with 5% CO2 in DMEM: F12 media (Life Technologies, Grand Island, NY) containing 100 units/mL penicillin/streptomycin (Life Technologies), 5% horse serum (Life Technologies), 20 ng/mL of epidermal growth factor (Life Technologies), 0.1 μg/mL cholera toxin (Sigma Aldrich, Saint Louis, MO), 10 μg/mL insulin (Sigma Aldrich, Saint Louis, MO), and 0.5 μg/mL hydrocortisone
3. Results
Protein levels of MPRα, MPRβ, MPRγ, MPRδ, MPRε, and PGRMC1 were determined in 15 μg total MCF10A and MCF12A cell lysate (Fig. 1). Additional positive and negative controls were used to assure the fidelity of our immunoblotting (data not shown). MPRα, MPRγ, MPRδ, MPRε, and PGRMC1 were all similarly produced in MCF10A and MCF12A cells, whereas MPRβ was expressed only in MCF10A cell lines. Notably, MCF10A and MCF12A cell lines are triple negative and therefore do not express PR.
To determine if
4. Discussion
Progestins are clearly involved in breast cancer biology, especially in light of the WHI and million women's studies demonstrating increased breast cancer risk in woman receiving CHT compared to EHT alone. In fact, women receiving EHT alone demonstrated a reduced risk (Rossouw et al., 2002, Anderson et al., 2004). Fundamentally, progestins induce biological response(s) by interacting with a receptor. Extensive study of the nuclear PR has provided a wealth of information. The recent discovery of
5. Conclusions
As noted, MCF10A and MCF12A cells are triple negative suggesting progestins effects occur through other receptors and not PR. Our data indicate two benign breast cell lines express several MPRs and PGRMC1 and respond differently to various types of progestins. The fact that MPA increased cell proliferation in MCF10A cells yet P4 did not underscores the importance of investigating the roles synthetic progestins play in breast cancer biology compared to natural P4. Concurrently, our data
Competing interests
The authors declare no competing interests.
Acknowledgements
The authors would like to thank Gail Silver, Nicole Sanchez, and Adriana Alire for assistance with conducting experiments. This research was funded by the Partnership for the Advancement of Cancer Research: NMSU/FHCRC, supported in part by NCI grants U54 CA132383 (NMSU) and U54 CA132381 (FHCRC); (awarded to A.K.A; awarded to R.L.A.), Cowboys for Cancer Research Grant (awarded to R.L.A.), the Howard Hughes Medical Institute Science Education Award #52006932 to NMSU, the Agriculture and Food
References (55)
- et al.
Breast cancer and hormone-replacement therapy: the million women study
Lancet
(2003) Progesterone receptor membrane component 1: an integrative review
J. Steroid Biochem. Mol. Biol.
(2007)- et al.
Proapoptotic effects of antiestrogens, progestins and androgen in breast cancer cells
J. Steroid Biochem. Mol. Biol.
(1999) - et al.
Comparison between steroid binding to membrane progesterone receptor alpha (mPRalpha) and to nuclear progesterone receptor: correlation with physicochemical properties assessed by comparative molecular field analysis and identification of mPRalpha-specific agonists
Steroids
(2010) - et al.
The effect of progesterone, testosterone and synthetic progestogens on growth factor- and estradiol-treated human cancerous and benign breast cells
Eur. J. Obstet. Gynecol. Reprod. Biol.
(2006) - et al.
Characterization of the stimulatory effect of medroxyprogesterone acetate and chlormadinone acetate on growth factor treated normal human breast epithelial cells
J. Steroid Biochem. Mol. Biol.
(2006) - et al.
Convergence of progesterone and epidermal growth factor signaling in breast cancer. Potentiation of mitogen-activated protein kinase pathways
J. Biol. Chem.
(1998) - et al.
Gene expression profiling changes induced by a novel Gemini vitamin D derivative during the progression of breast cancer
Biochem. Pharmacol.
(2006) - et al.
Progestin inhibition of cell death in human breast cancer cell lines
J. Steroid Biochem. Mol. Biol.
(2006) - et al.
Membrane 5alpha-pregnane-3.20-dione (5alphaP) receptors in MCF-7 and MCF-10A breast cancer cells are up-regulated by estradiol and 5alphaP and down-regulated by the progesterone metabolites, 3alpha-dihydroprogesterone and 20alpha-dihydroprogesterone, with associated changes in cell proliferation and detachment
J. Steroid Biochem. Mol. Biol.
(2005)
All progestins are not created equal
Steroids
Characteristics of membrane progestin receptor alpha (mPRalpha) and progesterone membrane receptor component 1 (PGMRC1) and their roles in mediating rapid progestin actions
Front. Neuroendocrinol.
Progesterone generates cancer stem cells through membrane progesterone receptor-triggered signaling in basal-like human mammary cells
Cancer Lett.
Plasma membrane receptors for the cancer-regulating progesterone metabolites, 5alpha-pregnane-3.20-dione and 3alpha-hydroxy-4-pregnen-20-one in MCF-7 breast cancer cells
Biochem. Biophys. Res. Commun.
Opposing actions of the progesterone metabolites, 5alpha-dihydroprogesterone (5alphaP) and 3alpha-dihydroprogesterone (3alphaHP) on mitosis, apoptosis, and expression of Bcl-2, Bax and p21 in human breast cell lines
J. Steroid Biochem. Mol. Biol.
Effects of conjugated equine estrogen in postmenopausal women with hysterectomy: the Women's Health Initiative randomized controlled trial
JAMA
Cloning and characterization of an ovine intracellular seven transmembrane receptor for progesterone that mediates calcium mobilization
Endocrinology
Progesterone stimulates mitochondrial activity with subsequent inhibition of apoptosis in MCF-10A benign breast epithelial cells
Am. J. Physiol. Endocrinol. Metab.
Expression of membrane progesterone receptors (mPR/PAQR) in ovarian cancer cells: implications for progesterone-induced signaling events
Horm. Cancer
Progestin regulation of cellular proliferation
Endocr. Rev.
Human chorionic gonadotropin increases serum progesterone, number of corpora lutea and angiogenic factors in pregnant sheep
Reproduction
MCF10AT: a model for the evolution of cancer from proliferative breast disease
Am. J. Pathol.
Signal-transducing protein phosphorylation cascades mediated by Ras/Rho proteins in the mammalian cell: the potential for multiplex signalling
Biochem. J.
Progestin signaling through mPRalpha in Atlantic croaker granulosa/theca cell cocultures and its involvement in progestin inhibition of apoptosis
Endocrinology
Membrane progesterone receptors (mPRs) mediate progestin induced antimorbidity in breast cancer cells and are expressed in human breast tumors
Horm. Cancer
Progesterone inhibits growth and induces apoptosis in breast cancer cells: inverse effects on Bcl-2 and p53
Ann. Clin. Lab. Sci.
Breast cancer risk in relation to different types of hormone replacement therapy in the E3N-EPIC cohort
Int. J. Cancer
Cited by (36)
Progestins and breast cancer hallmarks: The role of the ERK1/2 and JNK pathways in estrogen receptor positive breast cancer cells
2024, Journal of Steroid Biochemistry and Molecular BiologyProgesterone partially recovers placental glucose transporters in dexamethasone-induced intrauterine growth restriction
2022, Reproductive BioMedicine OnlineCitation Excerpt :The current results showed that progesterone treatment prevented the reduction in GLUT1 and GLUT3 expression seen with DEX treatment at both gene and protein levels in the labyrinth zone on 19dg. The effect of progesterone on preserving GLUT1 and GLUT3 expressions seen in the present study and improving vascularization as seen in other studies (He et al., 2006; Kim and Moley, 2009; Salazar et al., 2016) are believed to be the main reasons behind improving fetal body weight detected with progesterone treatment. In IUGR pregnancies, placental efficiency usually increases as a way of compensation to prevent fetal growth restriction (Ain et al., 2005; Coan et al., 2011; Dubova et al., 2013; Wyrwoll et al., 2009).
Loss of β-Arrestins or six Gα proteins in HEK293 cells caused Warburg effect and prevented progesterone-induced rapid proteasomal degradation of progesterone receptor membrane component 1
2021, Journal of Steroid Biochemistry and Molecular BiologyMicroRNA-181a suppresses norethisterone-promoted tumorigenesis of breast epithelial MCF10A cells through the PGRMC1/EGFR–PI3K/Akt/mTOR signaling pathway
2021, Translational OncologyCitation Excerpt :Although miR-181a expression is found in MCF-10A cells [14], the role of miR-181a and the effects of NET on breast epithelial cells remain poorly defined. Both PGRMC1 and EGFR are expressed in MCF10A cells [17]. We hypothesized that NET promotes the tumorigenesis of breast epithelial cells by upregulating the PGRMC1/EGFR signaling pathway.
Prolactin: A hormone with diverse functions from mammary gland development to cancer metastasis
2021, Seminars in Cell and Developmental BiologyCitation Excerpt :This study demonstrates that the cross-talk between two hormones is more complicated than previously thought, and likely different in breast cancer from the normal mammary epithelium. While STAT5 links PRL and PR in normal mammary development, the MAPK and AKT pathways are also activated in response to these hormones [124,125]. It is possible they are actually the primary points of cross-talk for prolactin and progesterone signaling or are of increased importance in the tumor setting.