Hepatocyte Growth Factor Is Required for Progestin-Induced Epithelial Cell Proliferation and Alveolar-Like Morphogenesis in Serum-Free Culture of Normal Mammary Epithelial Cells

Sunil, N

doi:10.1210/en.143.8.2953

Cited by 10 publications

(13 citation statements)

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“…The underlying mechanism of progesterone-induced side branch formation is not known. Based on the present results, we hypothesize that the progestin-induced blunting of HGF-induced tubulogenesis in vitro mimics progesterone-induced side branching in vivo and is likely mediated by similar mechanisms (13).…”

Section: Discussionsupporting

confidence: 63%

“…Cultures were maintained in basal medium (BM) [serumand phenol red-free DMEM/F12 (Sigma, St. Louis, MO), supplemented with 100 ng/ml human recombinant insulin, 1 mg/ml fatty acid-free BSA (fraction V), 100 g/ml penicillin, and 50 g/ml streptomycin] with or without or in various combinations of HGF (50 ng/ml; Sigma), promogestone (20 nM R5020 synthetic progestin; PerkinElmer, Boston, MA), mifepristone (100 nM RU486; Sigma), and 20 nM RANKL (200 ng/ml; R&D Systems, Minneapolis, MN). The synthetic progestin R5020 was used for culture studies because it is more stable and less rapidly metabolized than progesterone (13).…”

Section: Three-dimensional Collagen Culturesmentioning

confidence: 99%

“…This collagen gel culture model has been used extensively to study lung (9), salivary gland (10), and kidney (11) tubule development and mammary ductal development (12)(13)(14). Estrogen and progesterone receptors (PR) are well maintained in mammary organoids, and this culture model has proven useful to study steroid hormone action in mammary organoids (15).…”

mentioning

confidence: 99%

“…Estrogen alone or combined with progestin does not have an effect on proliferation or morphological changes (14). However, we and others have shown that primary mammary organoids composed of luminal epithelial cells (LEC) and myoepithelial cells (MEC) treated with growth factors, particularly hepatocyte growth factor (HGF) an estrogen-induced fibroblast-derived paracrine growth factor (14), develop tubules that resemble mammary ducts in vivo (12)(13)(14). In this culture model, we have defined the morphological steps during tubulogenesis (15).…”

mentioning

confidence: 99%

“…Notably, HGF-induced tubulogenesis is modulated by the addition of a synthetic progestin, R5020, resulting in shorter tubules (13,15). No extensions or tubules are formed in response to treatment with progestin alone, but instead, progestin-alone treatment causes central lumen formation within the body of the organoid by topographically localized apoptosis (13,15). The observed R5020-induced shortening of tubules induced by HGF has led us to hypothesize that this progestin-mediated effect in vitro is analogous to progesteronemediated side branching in vivo.…”

mentioning

confidence: 99%

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A Potential Role of Progestin-Induced Laminin-5/α6-Integrin Signaling in the Formation of Side Branches in the Mammary Gland

et al. 2012

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Mammary organoids from adult mice produce tubules, analogous to mammary ducts in vivo, in response to hepatocyte growth factor (HGF) when cultured in collagen gels. The combination of HGF plus progestin (R5020) causes reduced tubule number and length. We hypothesized that the inhibitory effect on tubulogenesis was due to progestin-mediated alteration of HGF/c-Met signaling. Using molecular inhibitors and short hairpin RNA, it was determined that HGF activation of Ras-related C3 botulinum toxin substrate (Rac1) was required for the formation of cytoplasmic extensions, the first step of tubulogenesis, and that Rac1 activity was Src kinase (Src) and focal adhesion kinase (FAK) dependent. The highly novel finding was that R5020 reduced tubulogenesis by up-regulating and increasing extracellular laminin and α6-integrin ligation to reduce activation of the Src, focal adhesion kinase, and Rac1 pathway. Receptor activator of nuclear factor-κB ligand, another progesterone-induced paracrine factor, did not replicate this effect of R5020. The inhibitory effect of R5020 on tubulogenesis was likely mediated through progesterone receptor (PR) isoform A (PRA), because PRA is the predominant PR isoform expressed in the organoids, and the progestin-induced effect was prevented by the PR antagonist RU486. These results provide a plausible mechanism that explains progestin/PRA-mediated blunting of HGF-induced tubulogenesis in vitro and is proposed to be relevant to progesterone/PRA-induced side-branching in vivo during pregnancy.

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Section: Discussionsupporting

confidence: 63%

Section: Three-dimensional Collagen Culturesmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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A Potential Role of Progestin-Induced Laminin-5/α6-Integrin Signaling in the Formation of Side Branches in the Mammary Gland

et al. 2012

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Dynamics of the Epithelium During Canalization of the Rat Ventral Prostate

Bruni‐Cardoso

Carvalho

2007

The Anatomical Record

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Outgrowth and branching of solid cords are the initial events in postnatal prostatic morphogenesis. These processes involve cell proliferation and their projection into the stroma and precede epithelial canalization. The purpose of the present study was to examine the dynamics of the prostate epithelium during canalization of the rat ventral prostate in the first week of postnatal development using histological, stereological, and ultrastructural analyses. The terminal deoxynucleotidyltransferase [TdT]-mediated deoxy-UTP nick end labeling assay was used to investigate the occurrence of DNA fragmentation. Our results demonstrate that canalization of the prostate epithelium starts as early as on day 1 (24 hr after birth) and progresses thereafter. By the end of the first week (day 6), luminal volume density reached 3% (P < 0.05) of the organ. Canalization was the result of epithelial cell differentiation and apoptosis. The former involved organization of the epithelial cells into a single layer sitting on the basement membrane, polarization, enlargement of secretory organelles and accumulation of secretory vesicles, microvilli formation, and establishment of the adult pattern of cell junctions. The latter was observed to occur mostly to epithelial cells not in contact with the basement membrane. Structures of variable electron density were observed in the developing lumen. In conclusion, different phenomena seem to be involved in the canalization of the rat ventral prostate. However, it was evident from the present results that complex epithelial cell fate decisions take place during this process.

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MiR-126-3p regulates progesterone receptors and involves development and lactation of mouse mammary gland

Cui

Qing-zhang

et al. 2011

Mol Cell Biochem

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MicroRNAs (miRNAs) are small (18-22 nucleotide) non-coding, endogenous regulatory RNA molecules, and they regulate gene expression at the post-transcriptional level through binding to their target mRNAs by base-pairing and subsequently inducing either translational repression or mRNA destabilization by plants, animals, and some viruses. In this study, combining microarray techniques with qRT-PCR, we found that miR-126-3p expression showed significant difference in the mouse mammary cycle during pregnancy, particularly on transition from pregnancy to lactation. Bioinformatics were used to predict target gene of miR-126-3p, and luciferase activity assay to test it, it showed that the progesterone receptor (PGR) 3'UTR is directly targeted by miR-126-3p. In this study, mouse mammary epithelial cells as cell model in vitro were used to study the function of miR-126-3p. Using gene silencing and over-expression for miR-126-3p, the expression of PGR protein and the secretion of casein were detected by western blotting and HPLC, respectively. To determine whether miR-126-3p can affect mouse mammary epithelial cells viability, cells were analyzed by CASY-YY. In conclusion, PGR gene confirmed miR-126-3p target genes through luciferase activity and western blotting. And miR-126-3p could also inhibit proliferation of mouse mammary epithelial cells (P < 0.01) and expression of β-casein (P < 0.01), and down-regulate PGR protein (P < 0.05). Our results suggested that miR-126-3p inhibited expression of PGR protein level as well as the proliferation of mammary epithelial cells, therefore miR-126-3p could play an important role in the process of mammary gland development.

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Hepatocyte Growth Factor Is Required for Progestin-Induced Epithelial Cell Proliferation and Alveolar-Like Morphogenesis in Serum-Free Culture of Normal Mammary Epithelial Cells

Cited by 10 publications

References 0 publications

A Potential Role of Progestin-Induced Laminin-5/α6-Integrin Signaling in the Formation of Side Branches in the Mammary Gland

A Potential Role of Progestin-Induced Laminin-5/α6-Integrin Signaling in the Formation of Side Branches in the Mammary Gland

Dynamics of the Epithelium During Canalization of the Rat Ventral Prostate

MiR-126-3p regulates progesterone receptors and involves development and lactation of mouse mammary gland

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