scholarly journals IGFBP-3 Induced by Ribotoxic Stress Traffics From the Endoplasmic Reticulum to the Nucleus in Mammary Epithelial Cells

2018 ◽  
Vol 3 (3) ◽  
pp. 517-536 ◽  
Author(s):  
Allyson Agostini-Dreyer ◽  
Amanda E Jetzt ◽  
Jennifer Skorupa ◽  
Jennifer Hanke ◽  
Wendie S Cohick
Keyword(s):  
Endoplasmic Reticulum ◽  
Epithelial Cells ◽  
Nuclear Localization ◽  
Nuclear Translocation ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Conditioned Media ◽  
Potent Inducer ◽  
Bovine Mammary Epithelial Cells ◽  
Igfbp 3

Abstract IGF-binding protein (IGFBP)-3 is a multifunctional protein that can exert IGF-independent effects on apoptosis. Anisomycin (ANS) is a potent inducer of IGFBP-3 production in bovine mammary epithelial cells (MECs), and knockdown of IGFBP-3 attenuates ANS-induced apoptosis. IGFBP-3 is present in the nucleus and the conditioned media in response to ANS. The goal of this study was to determine whether ribotoxic stress induced by ANS or a second ribotoxin, deoxynivalenol (DON), specifically regulates transport of IGFBP-3 to the nucleus and to determine the pathway by which it traffics. In ribotoxin-treated cells, both endogenous IGFBP-3 and transfected IGFBP-3 translocated to the nucleus. Inhibition of the nuclear transport protein importin-β with importazole reduced ribotoxin-induced nuclear IGFBP-3. Immunoprecipitation studies showed that ANS induced the association of IGFBP-3 and importin-β, indicating that ribotoxins specifically induce nuclear translocation via an importin-β‒dependent mechanism. To determine whether secretion of IGFBP-3 is required for nuclear localization, cells were treated with Pitstop 2 or brefeldin A to inhibit clathrin-mediated endocytosis or overall protein secretion, respectively. Neither inhibitor affected nuclear localization of IGFBP-3. Although the IGFBP-3 present in both the nucleus and conditioned media was glycosylated, secreted IGFBP-3 exhibited a higher molecular weight. Deglycosylation experiments with endoglycosidase Hf and PNGase indicated that secreted IGFBP-3 completed transit through the Golgi apparatus, whereas intracellular IGFBP-3 exited from the endoplasmic reticulum before transit through the Golgi. In summary, ANS and DON specifically induced nuclear localization of nonsecreted IGFBP-3 via an importin-β‒mediated event, which may play a role in their ability to induce apoptosis in MECs.

scholarly journals Receptor Activator of NF-κB Ligand Regulates the Proliferation of Mammary Epithelial Cells via Id2

2006 ◽  
Vol 26 (3) ◽  
pp. 1002-1013 ◽  
Author(s):  
Nam-Shik Kim ◽  
Hyun-Ju Kim ◽  
Bon-Kyoung Koo ◽  
Min-Chul Kwon ◽  
Young-Woong Kim ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Epithelial Cells ◽  
Nuclear Localization ◽  
Nuclear Translocation ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
In Vivo Studies ◽  
Nuclear Localization Sequence ◽  
Receptor Activator ◽  
Rankl Stimulation

ABSTRACT Receptor activator of NF-κB ligand (RANKL) is a key regulator for mammary gland development during pregnancy. RANKL-deficient mice display impaired development of lobulo-alveolar mammary structures. Similar mammary gland defects have been reported in mice lacking Id2. Here we report that RANKL induces the proliferation of mammary epithelial cells via Id2. RANKL triggers marked nuclear translocation of Id2 in mammary epithelial cells. In vivo studies further demonstrated the defective nuclear translocation of Id2, but the normal expression of cyclin D1, in the mammary epithelial cells of rankl −/− mice. In vitro studies with nuclear localization sequence-tagged Id2 revealed that the nuclear localization of Id2 itself is critical for the downregulation of p21 promoter activity. Moreover, RANKL stimulation failed to induce cell growth and to downregulate p21 expression in Id2 −/− mammary epithelial cells. Our results indicate that the inhibitor of helix-loop-helix protein, Id2, is critical to control the proliferation of mammary epithelial cells in response to RANKL stimulation.

scholarly journals Metformin Inhibits Lipoteichoic Acid–Induced Oxidative Stress and Inflammation Through AMPK/NRF2/NF-κB Signaling Pathway in Bovine Mammary Epithelial Cells

2021 ◽  
Vol 8 ◽  
Author(s):  
Abdelaziz Adam Idriss Arbab ◽  
Xubin Lu ◽  
Ismail Mohamed Abdalla ◽  
Amer Adam Idris ◽  
Zhi Chen ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Epithelial Cells ◽  
Signaling Pathway ◽  
Nuclear Localization ◽  
Mammary Epithelial Cells ◽  
Lipoteichoic Acid ◽  
Mammary Epithelial ◽  
Heme Oxygenase 1 ◽  
Nuclear Factor ◽  
Bovine Mammary Epithelial Cells

The objective of this research was to explore the effect of metformin on the lipoteichoic acid (LTA)–induced mastitis model using isolated primary bovine mammary epithelial cells (PBMECs). The PBMECs were exposed to either 3 mM metformin for 12 h as a metformin group (MET) or 100 μg/mL LTA for 6 h as LTA group (LTA). Cells pretreated with 3 mM metformin for 12 h followed by washing and 100 μg/mL LTA exposure for 6 h served as the MET + LTA group. Phosphate-buffered saline was added to cells as the control group. PBMECs pretreated with different metformin doses were analyzed by a flow cytometry (annexin V–fluorescein isothiocyanate assay) to detect the cell apoptotic rate. We performed quantitative reverse transcriptase–polymerase chain reaction and Western blot analysis to evaluate the inflammatory and oxidative responses to metformin and LTA by measuring cellular cytotoxicity, mRNA expression, and protein expression. Immunofluorescence was used to evaluate nuclear localization. The results showed that the gene expression of COX2, IL-1β, and IL-6 significantly increased in the cells challenged with LTA doses compared to control cells. In inflammatory PBMECs, metformin attenuated LTA-induced expression of inflammatory genes nuclear factor κB (NF-κB) p65, tumor necrosis factor α, cyclooxygenase 2, and interleukin 1β, as well as the nuclear localization and phosphorylation of NF-κBp65 protein, but increased the transcription of nuclear factor erythroid 2–related factor 2 (Nrf2) and Nrf2-targeted antioxidative genes heme oxygenase-1 (HO-1) and Gpx1, as well as the nuclear localization of HO-1 protein. Importantly, metformin-induced activation of Nrf2 is AMP-activated protein kinase (AMPK)–dependent; as metformin-pretreated PBMECs activated AMPK signaling via the upregulation of phosphorylated AMPK levels, cell pretreatment with metformin also reversed the translocation of Nrf2 that was LTA inhibited. This convergence between AMPK and Nrf2 pathways is essential for the anti-inflammatory effect of metformin in LTA-stimulated PBMECs. Altogether, our results indicate that metformin exerts anti-inflammation and oxidative stress through regulation of AMPK/Nrf2/NF-κB signaling pathway, which highlights the role of AMPK as a potential therapeutic strategy for treatment of bovine mastitis.

scholarly journals Zearalenone induces apoptosis in bovine mammary epithelial cells by activating endoplasmic reticulum stress

2019 ◽  
Vol 102 (11) ◽  
pp. 10543-10553 ◽  
Author(s):  
YuRong Fu ◽  
YongCheng Jin ◽  
Yun Zhao ◽  
AnShan Shan ◽  
HengTong Fang ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Epithelial Cells ◽  
Endoplasmic Reticulum Stress ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Bovine Mammary Epithelial Cells

scholarly journals Effects of fatty acids on inducing endoplasmic reticulum stress in bovine mammary epithelial cells

2020 ◽  
Vol 103 (9) ◽  
pp. 8643-8654
Author(s):  
Mst Mamuna Sharmin ◽  
Moeko Mizusawa ◽  
Satoko Hayashi ◽  
Wataru Arai ◽  
Shotaro Sakata ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Endoplasmic Reticulum ◽  
Epithelial Cells ◽  
Endoplasmic Reticulum Stress ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Bovine Mammary Epithelial Cells

scholarly journals IGF-1 induces IRE1-XBP1–dependent endoplasmic reticulum biogenesis in bovine mammary epithelial cells

2021 ◽  
Author(s):  
Mst Mamuna Sharmin ◽  
Satoko Hayashi ◽  
Makoto Miyaji ◽  
Hiroshi Ishizaki ◽  
Hiroki Matsuyama ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Epithelial Cells ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Bovine Mammary Epithelial Cells

scholarly journals Polydatin Protects Bovine Mammary Epithelial Cells against Zearalenone-Induced Apoptosis by Inhibiting Oxidative Responses and Endoplasmic Reticulum Stress

Toxins ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 121
Author(s):  
Yurong Fu ◽  
Yongcheng Jin ◽  
Anshan Shan ◽  
Jing Zhang ◽  
Hongyu Tang ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Treatment Group ◽  
Epithelial Cells ◽  
Er Stress ◽  
Cell Apoptosis ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Bovine Mammary Epithelial Cells ◽  
Stress Related Genes ◽  
Induced Apoptosis

Zearalenone (ZEA) is a mycotoxin of the Fusarium genus that can cause endoplasmic reticulum (ER) stress and Apoptosis in bovine mammary epithelial cells (MAC-T). Polydatin (PD), a glycoside purified from Polygonum cuspidatum, has antioxidant properties. This study aimed to explore whether PD can alleviate ZEA-induced damage on bovine mammary epithelial cells (MAC-T). We found that incasing the concentration of ZEA (0, 7.5, 15, 30, 60, 90, 120, and 240 μM) gradually decreased the cell viability. PD treatment alone at 5, 10, and 20 μM did not affect cell viability. Follow-up studies then applied 30 μM of ZEA and 5 μM of PD to treat cells; the results showed that the ZEA + PD treatment group effectively reduced cell oxidative damage compared with the ZEA treatment group. The qPCR analysis showed that ZEA treatment significantly up-regulated the expression of ER stress-related genes, relative to the control. However, adding PD significantly down-regulated the expression of ER stress-related genes. The cell apoptosis detection results showed that, compared with the ZEA treatment group, the ZEA + PD treatment group down-regulated the Bax gene and up-regulated the Bcl-2 gene expressions, which reduced the cell apoptosis rate and Caspase-3 activity. Taken together, these results indicate that PD reduces ZEA-induced apoptosis by inhibiting oxidative damage and ER stress.

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