A model of secreting murine mammary epithelial HC11 cells comprising endogenous Bcrp/Abcg2 expression and function

Tallkvist, Jonas; Yagdiran, Yagmur; Danielsson, Louise; Oskarsson, Agneta

doi:10.1007/s10565-015-9298-5

Cited by 6 publications

(9 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Following this EGF depletion step differentiation of the cells was accomplished by culturing for an additional 72 hours in serum- and EGF- free medium containing 1 mg/L prolactin and 1μM hydrocortisone. Differentiation of the cells was assessed by measuring induction of β-casein (Csn2) gene expression as well as examination of cellular morphology as described previously [39]. …”

Section: Methodsmentioning

confidence: 99%

“…We have previously characterized gene expressions of transporters in mammary gland of mice at different lactation stages and in HC11 cells. Gene expressions of Csn2 (β-casein) , Bcrp , Mdr1 , Mrp1 , Oatp1a5 , Octn1 and Oct1 were altered during gestation and lactation in mice mammary glands and in HC11 cells the expression patterns were affected by differentiation [9, 39]. …”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Staphylococcus aureus and Lipopolysaccharide Modulate Gene Expressions of Drug Transporters in Mouse Mammary Epithelial Cells Correlation to Inflammatory Biomarkers

et al. 2016

Self Cite

View full text Add to dashboard Cite

Inflammation in the mammary gland (mastitis) is the most common disease in dairy herds worldwide, often caused by the pathogens Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). Little is known about the effects of mastitis on drug transporters and the impact on transporter-mediated excretion of drugs into milk. We used murine mammary epithelial HC11 cells, after lactogenic differentiation into a secreting phenotype, and studied gene expressions of ABC- and SLC- transporters after treatment of cells with S. aureus and lipopolysaccharide, an endotoxin secreted by E. coli. The studied transporters were Bcrp, Mdr1, Mrp1, Oatp1a5, Octn1 and Oct1. In addition, Csn2, the gene encoding β-casein, was analyzed. As biomarkers of the inflammatory response, gene expressions of the cytokines Il6 and Tnfα and the chemokine Cxcl2 were determined. Our results show that S. aureus and LPS treatment of cells, at non-cytotoxic concentrations, induced an up-regulation of Mdr1 and of the inflammatory biomarkers, except that Tnfα was not affected by lipopolysaccharide. By simple regression analysis we could demonstrate statistically significant positive correlations between each of the transporters with each of the inflammatory biomarkers in cells treated with S. aureus. The coefficients of determination (R2) were 0.7–0.9 for all but one correlation. After treatment of cells with lipopolysaccharide, statistically significant correlations were only found between Mdr1 and the two parameters Cxcl2 and Il6. The expression of Csn2 was up-regulated in cells treated with S. aureus, indicating that the secretory function of the cells was not impaired. The strong correlation in gene expressions between transporters and inflammatory biomarkers may suggest a co-regulation and that the transporters have a role in the transport of cytokines and chemokines. Our results demonstrate that transporters in mammary cells can be affected by infection, which may have an impact on transport of essential compounds and contaminants into milk.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Staphylococcus aureus and Lipopolysaccharide Modulate Gene Expressions of Drug Transporters in Mouse Mammary Epithelial Cells Correlation to Inflammatory Biomarkers

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

“…However, some of these transporters feature broad substrate specificities and have been demonstrated to mediate active transport also of drugs and toxic chemicals into milk [ 7 , 9 , 14 – 16 ]. Transporters with broad substrate specificities and altered expressions in lactating as compared to resting mammary epithelial cells include breast cancer resistance protein (BCRP/ABCG2) [ 6 , 7 , 13 , 17 , 18 ], multidrug resistance protein 1(MDR1/ABCB1) [ 7 , 18 ], multidrug resistance-associated protein 1 (MRP1/ABCC1) [ 7 , 10 ], solute carrier organic anion transporter family member 1A2 (OATP1A2/SLCO1A2), organic cation transporter novel protein type 1 (OCTN1/SLC22A4) [ 10 ] and organic cation transporter 1 (OCT1/SLC22A1) [ 16 ]…”

Section: Introductionmentioning

confidence: 99%

“…After treatment the cells feature a secreting phenotype [ 25 – 27 ]. We have recently demonstrated that BCRP and MDR1 are expressed in HC11 cells and are up- and downregulated, respectively by differentiation of the cells, similar to the situation in vivo during lactation [ 18 ]. In the present study we also used bovine mammary epithelial BME-UV cells, derived from a lactating Holstein cow, and previously reported to express MDR1 [ 28 ].…”

Section: Introductionmentioning

confidence: 99%

ABC- and SLC-Transporters in Murine and Bovine Mammary Epithelium - Effects of Prochloraz

et al. 2016

Self Cite

View full text Add to dashboard Cite

Some chemicals are ligands to efflux transporters which may result in high concentrations in milk. Limited knowledge is available on the influence of maternal exposure to chemicals on the expression and function of transporters in the lactating mammary gland. We determined gene expression of ABC and SLC transporters in murine mammary tissue of different gestation and lactation stages, in murine mammary cells (HC11) featuring resting and secreting phenotypes and in bovine mammary tissue and cells (BME-UV). Effects on transporter expression and function of the imidazole fungicide prochloraz, previously reported to influence BCRP in mammary cells, was investigated on transporter expression and function in the two cell lines. Transporters studied were BCRP, MDR1, MRP1, OATP1A5/OATP1A2, OCTN1 and OCT1. Gene expressions of BCRP and OCT1 in murine mammary glands were increased during gestation and lactation, whereas MDR1, MRP1, OATP1A5 and OCTN1 were decreased, compared to expressions in virgins. All transporters measured in mammary glands of mice were detected in bovine mammary tissue and in HC11 cells, while only MDR1 and MRP1 were detected in BME-UV cells. Prochloraz treatment induced MDR1 gene and protein expression in both differentiated HC11 and BME-UV cells and increased protein function in HC11 cells, resulting in decreased accumulation of the MDR1 substrate digoxin. In conclusion, our results demonstrate that murine (HC11) and bovine (BME-UV) mammary epithelial cells can be applied to characterize expression and function of transporters as well as effects of contaminants on the mammary transporters. An altered expression, induced by a drug or toxic chemical, on any of the transporters expressed in the mammary epithelial cells during lactation may modulate the well-balanced composition of nutrients and/or secretion of contaminants in milk with potential adverse effects on breast-fed infants and dairy consumers.

show abstract

“…The same group also demonstrated protoporphyria in these animals, indicating a role of ABCG2 in hematopoiesis, possibly through interaction with protoporphyrin IX, which was supported by further studies. Moreover, the functional relevance of ABCG2 was demonstrated in other mammals, being responsible for the accumulation of chemicals in milk [11], or being an important protector of retinal cells against toxic agents in domestics cats [12]. In human populations, certain SNPs in the ABCG2 gene have been linked to the occurrence of gout [13].…”

Section: Introductionmentioning

confidence: 99%

Functional characterization of the ABCG2 5′ non-coding exon variants: Stem cell specificity, translation efficiency and the influence of drug selection

Sándor

Jordanidisz

Schamberger

et al. 2016

Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanism

View full text Add to dashboard Cite

Functional characterization of the ABCG2 5' non-coding exon variants: stem cell specificity, translation efficiency and the influence of drug selection, BBA -Gene Regulatory Mechanisms (2016Mechanisms ( ), doi: 10.1016Mechanisms ( /j.bbagrm.2016 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT2 Abstract ABCG2 is a multidrug transporter with wide substrate specificity, and is believed to protect several cell types from various xenobiotics and endobiotics. This "guardian" function is important in numerous cell types and tissue barriers but becomes disadvantageous by being responsible for the multidrug resistance phenotype in certain tumor cells. ABCG2 regulation at the protein level has already been extensively studied, however, regulation at the mRNA level, especially the functional role of the various 5' untranslated exon variants (5' UTRs) has been elusive. In the present work, we describe a comprehensive characterization of four ABCG2 mRNA variants with different exon 1 sequences, investigate drug inducibility, stem cell specificity, mRNA stability, and translation efficiency. Although certain variants (E1B and E1C) are considered as "constitutive" mRNA isoforms, we show that chemotoxic drugs significantly alter the expression pattern of distinct ABCG2 mRNA isoforms. When examining human embryonic stem cell lines, we provide evidence that variant E1A has an expression pattern coupled to undifferentiated stem cell stage, as its transcript level is regulated parallel to mRNAs of Oct4 and Nanog pluripotency marker genes. When characterizing the four exon 1 variants we found no significant differences in terms of mRNA stabilities and half-lives of the isoforms. In contrast, variant E1U showed markedly lower translation efficiency both at the total protein level or regarding the functional presence in the plasma membrane. Taken together, these results indicate that the different 5' UTR variants play an important role in cell type specific regulation and fine tuning of ABCG2 expression.

show abstract

A model of secreting murine mammary epithelial HC11 cells comprising endogenous Bcrp/Abcg2 expression and function

Cited by 6 publications

References 43 publications

Staphylococcus aureus and Lipopolysaccharide Modulate Gene Expressions of Drug Transporters in Mouse Mammary Epithelial Cells Correlation to Inflammatory Biomarkers

Staphylococcus aureus and Lipopolysaccharide Modulate Gene Expressions of Drug Transporters in Mouse Mammary Epithelial Cells Correlation to Inflammatory Biomarkers

ABC- and SLC-Transporters in Murine and Bovine Mammary Epithelium - Effects of Prochloraz

Functional characterization of the ABCG2 5′ non-coding exon variants: Stem cell specificity, translation efficiency and the influence of drug selection

Contact Info

Product

Resources

About