All-trans retinoic acid induced differentiation of rat mammary epithelial cells cultured in serum-free medium

1998 ◽  
Vol 21 (3) ◽  
pp. 298-304 ◽  
Author(s):  
Min Hyo Ki ◽  
Kee-Joo Paik ◽  
Ji Hyeon Lee ◽  
Hae Young Chung ◽  
Kyung Hee Lee ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Epithelial Cells ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Trans Retinoic Acid ◽  
All Trans Retinoic Acid ◽  
Cells Cultured

Growth and differentation of rat mammary epithelial cells cultured in serum-free medium

1997 ◽  
Vol 20 (4) ◽  
pp. 297-305 ◽  
Author(s):  
Dong Yeum Kim ◽  
Byung-Hak Jhun ◽  
Kyung Hee Lee ◽  
Seung Chul Hong ◽  
Kelly H. Clifton ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Cells Cultured

scholarly journals Protection of bovine mammary epithelial cells from hydrogen peroxide-induced oxidative cell damage by selenium

2018 ◽  
Vol 63 (No. 3) ◽  
pp. 94-102
Author(s):  
Y.M. Guo ◽  
J. Gong ◽  
Y.G. Zheng ◽  
B.L. Shi ◽  
X.Y. Guo ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Epithelial Cells ◽  
Oxidative Damage ◽  
Mammary Epithelial Cells ◽  
Mapk Pathway ◽  
Mammary Epithelial ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Bovine Mammary Epithelial Cells

The uncontrolled release of arachidonic acid (ARA) and its metabolism by lipoxygenase (LOX) pathway can induce and aggravate cellular oxidative stress. Selenium (Se) is an integral part of some antioxidative selenoproteins and may protect cells from oxidative damage by modulating ARA release and metabolism. The present study aimed to investigate the protective response of Se against hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>)-induced oxidative damage in bovine mammary epithelial cells (BMECs). The BMECs were incubated for 24 h in serum-free medium and then divided into four groups randomly. The cells in groups 1 and 2 were subsequently incubated for 30 h in serum-free medium containing 0 (control) and 50 nM Se (Se treatment group). The cells in groups 3 and 4 were incubated for 24 h in serum-free medium containing 0 and 50 nM Se, and then treated with 600 μM H<sub>2</sub>O<sub>2</sub> for 6 h (H<sub>2</sub>O<sub>2</sub> damage group and Se prevention group). The results showed that Se attenuated the H<sub>2</sub>O<sub>2</sub>-induced production of reactive oxygen species and the decrease of antioxidative enzymes as glutathione peroxidase (GPX), thioredoxin reductase (TrxR), selenoprotein P (SelP), superoxide dismutase, and catalase in BMECs. The preventive effects of Se on the decrease of selenoprotein activity were demonstrated further by the increase of mRNA expression for GPX1, TrxR1, and SelP, and protein expression for GPX1 and TrxR1. Pretreatment of cells with Se inhibited the H<sub>2</sub>O<sub>2</sub>-induced increase of mRNA expressions and activities for cytosolic phospholipase A2 and 5-lipoxygenase, ARA release, and 15-hydroperoxyeicosatetraenoic acid production. Se also blocked the H<sub>2</sub>O<sub>2</sub>-induced activation of p38 mitogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase but not that of extracellular signal-regulated kinase. These results suggested that Se may protect BMECs against H<sub>2</sub>O<sub>2</sub>-induced oxidative damage by increasing selenoproteins synthesis, inhibiting MAPK pathway, and then decreasing ARA release and its metabolism by LOX pathway.

Metabolism of 7,12-dimethylbenz[a]anthracene by mouse mammary epithelial cells cultured serum-free inside collagen gels

1988 ◽  
Vol 40 (2) ◽  
pp. 123-132 ◽  
Author(s):  
Raphael C. Guzman ◽  
Rebecca C. Osborn ◽  
Jack C. Bartley ◽  
Satyabrata Nandi
Keyword(s):  
Epithelial Cells ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Collagen Gels ◽  
Serum Free ◽  
Cells Cultured

Insulin/IGF1 Pathway Inhibition in Combination with All-Trans Retinoic Acid in Acute Promyelocytic Leukemia

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 1347-1347
Author(s):  
Tiphanie Durfort ◽  
Luca Mazzarella ◽  
Pier Giuseppe Pelicci
Keyword(s):  
Insulin Receptor ◽  
Drug Target ◽  
Promyelocytic Leukemia ◽  
Serum Free Medium ◽  
Free Medium ◽  
Atra Treatment ◽  
Serum Free ◽  
Trans Retinoic Acid ◽  
All Trans Retinoic Acid

Abstract Abstract 1347 Chemotherapy-free regimens are now a realistic goal in Acute Promyelocytic Leukemia (APL), as demonstrated by recent studies combining All-Trans Retinoic Acid (ATRA) with Arsenic Trioxide and Gemtuzumab Ozogamicin. Combinations of ATRA with kinase inhibitors, which have shown tremendous efficacy in several malignancies, are interesting alternatives but remain poorly characterized in APL. The Insulin-like growth factor 1 (IGF1) pathway is an attractive drug target because of its pervasive involvement in cell proliferation and metabolism of cancer cells. A variety of strategies to target the Insulin/IGF1 axis have been developed, but trials have struggled to demonstrate real efficacy over standard treatments, probably due to our still incomplete understanding of the intricacies of the downstream signalling cascade. Importantly, recent evidence suggests that inhibition of the sole IGF1 receptor may be inadequate because the structurally similar insulin receptor (IR) or IGF1R/IR heterodimeric receptors may be sufficient to support tumour growth. We dissected the IGF1R/IR pathway in the well established APL cell line NB4. IGF1 and insulin, but not the structurally similar IGF1R ligand IGF2, significantly increased NB4 growth in serum-free medium. ATRA treatment resulted in the downregulation, at both transcript and protein level, of several components of the proximal IR/IGF1R pathway: IGF1R (down by 64,9 ±1,7%), IR (down by 82,5 ±4,9%), the transducer proteins insulin receptor substrate 1 (IRS1) (down by 85,7 %) and IRS2 insulin receptor IRS2) by 98,1% (+/− 0,8). However, the downstream cascade remained ligand-responsive: stimulation of serum-starved cells with IGF1 induced a dose-dependent phosphorylation of AKT, FOXO, the mTOR target ribosomal protein S6 and ERK1-2. In addition, baseline levels of phospho-ERK were elevated after ATRA treatment. This suggested that ATRA-treated cells may increase their dependence on IGF1R for their survival. In agreement with this model, NB4 cell growth was completely inhibited by the anti-IGF1R antibody aIR3, but concomitant treatment with ATRA halved the half-maximal dose (IC50) of aIR3 to 1 ug/ml and promoted apoptosis (as assessed by flow cytometry). To identify IGF1R-targeting compounds with higher efficacy we screened novel small molecule inhibitors and found that the imidazopyrazine-derivative OSI-906 stopped NB4 cell growth with an IC50 of 1.5 μM in IGF1-supplemented serum-free medium. This compound is an orally available dual IR/IGF1R inhibitor, currently on trial for several solid tumours. In conclusion, we show that the Insulin/IGF1 signalling pathway is modulated at different levels by ATRA in APL cell lines and that this pathway represents a suitable and attractive drug target in combination with ATRA treatment. On the basis of data presented here we are currently testing the in vivo efficacy of ATRA/OSI906 combinations in in vivo models of APL. Disclosures: Off Label Use: Linsitinib (OSI-906) is a potent, selective orally active inhibitor of the insulin-like growth factor-1 receptor (IGF-1R).

Lactoferrin expression in mammary epithelial cells is mediated by changes in cell shape and actin cytoskeleton

1997 ◽  
Vol 110 (22) ◽  
pp. 2861-2871 ◽  
Author(s):  
M.J. Close ◽  
A.R. Howlett ◽  
C.D. Roskelley ◽  
P.Y. Desprez ◽  
N. Bailey ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Mammary Gland ◽  
Cell Shape ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Physical Constraints ◽  
Beta Casein

Lactoferrin is a secreted iron binding protein which is expressed during normal functional development of mammary epithelium. Murine mammary epithelial cell lines competent for milk protein expression were used to identify microenvironmental factors that regulate lactoferrin expression. While lactoferrin was not expressed in adherent monolayer cultures under standard subconfluent conditions on plastic, lactoferrin mRNA and protein steadily accumulated when the cells aggregated to form spheroids on a reconstituted basement membrane gel. However, unlike other milk proteins such as beta-casein, lactoferrin expression was also induced at high cell density in the absence of exogenously added basement membrane or prolactin. These results led us to examine whether changes in cell growth, cell-cell interactions and/or cell shape were responsible for regulation of lactoferrin gene expression. Rounded, non-proliferating cells in suspension in serum-free medium expressed lactoferrin even as single cells. Conversely, lactoferrin expression could be inhibited in non-proliferative cells in serum-free medium by maintaining them in contact with an air-dried extracellular matrix which caused the cells to retain flat, spread morphologies. These findings indicated that cessation of cell growth was not sufficient, that cell-cell interactions were not required, and that cell culture conditions which minimize cell spreading may be important in maintaining lactoferrin expression. Additional data supporting this latter concept were generated by treating spread cells with cytochalasin D. The resulting disruption of microfilament assembly induced both cell rounding and lactoferrin expression. Shape-dependent regulation of lactoferrin mRNA was both transcriptional and post-transcriptional. Surprisingly, treatment of rounded cells with a transcription inhibitor, actinomycin D, produced a stabilization of lactoferrin mRNA, suggesting that transcription of an unstable factor is required for degradation of lactoferrin mRNA. Importantly, lactoferrin mRNA expression was regulated similarly in early passage normal human mammary epithelial cells. In vivo, the changing extracellular matrix components of the mammary gland during different stages of normal and abnormal growth and differentiation may provide different physical constraints on the configurations of cell surface molecules. These physical constraints may be communicated to the cell interior through mechanical changes in the cytoskeleton. Unlike beta-casein whose expression is upregulated by specific integrin-mediated signals, lactoferrin may be representative of a class of proteins synthesized in the mammary gland using basal transcriptional and translational machinery. The suppression of lactoferrin expression that is observed in monolayer culture and in malignant tissues may reflect inappropriate cell shapes and cytoskeletal structures that are manifested under these conditions.

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