Gene Expression Profiling of Systems Involved in the Metabolism and the Disposition of Xenobiotics: Comparison between Human Intestinal Biopsy Samples and Colon Cell Lines

Bourgine, Joanna; Billaut-Laden, Ingrid; Happillon, M.; Lo-Guidice, Jean-Marc; Maunoury, V.; Imbenotte, M.; Broly, Franck

doi:10.1124/dmd.111.042465

Cited by 66 publications

(59 citation statements)

References 38 publications

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“…Nucleosides and their analog drugs generally have hydrophilic properties and diffuse slowly across the cell membrane; therefore, nucleoside transporters expressed in the small intestine largely contribute to their oral absorption (Endres et al, 2009;Okayama et al, 2012;Ishida et al, 2013). Previous studies reported that Caco-2 cells did not express CNT1, CNT2, or CNT3 (Ward and Tse, 1999;Bourgine et al, 2012), which was consistent with the very low expression of CNTs observed in our study (Fig. 3B).…”

Section: Discussionsupporting

confidence: 92%

“…3) have been well documented in previous studies (Schmiedlin-Ren et al, 1997;Sun et al, 2002;Bourgine et al, 2012). In this study, the expression profile of mRNA for the P450/UGT isoforms in HIECs was similar to that in Caco-2 cells, except for CYP2C9, CYP2C19, UGT1A6, and UGT2B7.…”

Section: Discussionsupporting

confidence: 87%

“…In this study, the expression profile of mRNA for the P450/UGT isoforms in HIECs was similar to that in Caco-2 cells, except for CYP2C9, CYP2C19, UGT1A6, and UGT2B7. Although the causes of these exceptions are not known, the differences of origins (normal versus tumoral tissue) between HIECs and Caco-2 cells (Bourgine et al, 2012) and medium additives, such as insulin (Martínez-Jiménez et al, 2006) and dexamethasone (Jemnitz et al, 2002), may contribute these observations. Among the mRNAs whose expression was examined, the mRNA expression (Fig.…”

Section: Discussionmentioning

confidence: 99%

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Human Small Intestinal Epithelial Cells Differentiated from Adult Intestinal Stem Cells as a Novel System for Predicting Oral Drug Absorption in Humans

et al. 2014

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Adult intestinal stem cells (ISCs) possess both a long-term proliferation ability and differentiation capability into enterocytes. As a novel in vitro system for the evaluation of drug absorption, we characterized a human small intestinal epithelial cell (HIEC) monolayer that differentiated from adult ISCs. Continuous proliferation/ differentiation from ISCs consistently conferred the capability of maturation of enterocytes to HIECs over 25 passages. The morphologically matured HIEC monolayer consisted of polarized columnar epithelia with dense microvilli, tight junctions, and desmosomes 8 days after seeding onto culture inserts. Transepithelial electrical resistance across the monolayer was 9-fold lower in HIECs (98.9 V 3 cm 2 ) than in Caco-2 cells (900 V 3 cm 2 ), which indicated that the looseness of the tight junctions in the HIEC monolayer was similar to that in the human small intestine (approximately 40 V 3 cm 2 ). No significant differences were observed in the overall gene expression patterns of the major drug-metabolizing enzymes and transporters between the HIEC and Caco-2 cell monolayers. Furthermore, the functions of P-glycoprotein and breast cancer resistance protein in the HIEC monolayer were confirmed by the vectorial transport of marker substrates and their disappearance in the presence of specific inhibitors. The apparent drug permeability values of paracellularly transported compounds (fluorescein isothiocyanate-dextran 4000, atenolol, and terbutaline) and nucleoside transporter substrates (didanosine, ribavirin, and doxifluridine) in the HIEC monolayer were markedly higher than those of Caco-2 cells, whereas transcellularly transported drugs (pindolol and midazolam) were equally well permeated. In conclusion, the HIEC monolayer can serve as a novel and superior alternative to the conventional Caco-2 cell monolayer for predicting oral absorption in humans.

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Section: Discussionsupporting

confidence: 92%

Section: Discussionsupporting

confidence: 87%

Section: Discussionmentioning

confidence: 99%

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Human Small Intestinal Epithelial Cells Differentiated from Adult Intestinal Stem Cells as a Novel System for Predicting Oral Drug Absorption in Humans

et al. 2014

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“…Nevertheless, several proteins, including transporters, were over-or underexpressed. Bourgine et al ( 2012 ) compared the expression of 377 genes in HT29 and other intestinal cell lines that are used as in vitro models of the epithelium with the corresponding tissue biopsy. The results showed that differentiated HT29 cells and human colonic tissues do not appear to be signifi cantly different.…”

Section: Relevance To Human In Vivo Situationmentioning

confidence: 99%

HT29 Cell Line

Martínez-Maqueda

Miralles

Recio

2015

The Impact of Food Bioactives on Health

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The human colon adenocarcinoma cell line HT29, is not only used to study the biology of human colon cancers, but it is receiving special interest in studies focused on food digestion and bioavailability due to the ability to express characteristics of mature intestinal cells. In the differentiated phenotype, they are able to form a monolayer with tight junctions between cells and a typical apical brush border. In addition, these differentiated cells express brush-border-associated hydrolases typical of the small intestine although the enzymatic activity is lower than that found in vivo. Although they represent a valuable model due to their similarities with enterocytes of the small intestine, their limitations and the relevance to the in vivo situation are also considered in this chapter. The application of this cell line to transport studies of drugs and food compounds is illustrated, especially when the effect of the mucus layer is considered or used as co-culture in combination with Caco-2 cells. They have also been frequently used to study the intestinal immune response to bacterial infection, and microorganism survival, adhesion or invasion. Finally, the use of these cells to evaluate the effect of several food compounds and mucin secretion is summarized. Keywords HT29 • Cell differentiation • Transport studies • Intestinal cell OriginThe human colon adenocarcinoma cell line HT29 was isolated from a primary tumor of a 44 years old Caucasian female in 1964 by Fogh and Trempe ( 1975 ). Since then, many cell lines have been derived from human colon cancers. Initially, this cell line was used to study different aspects of the biology of human cancers. However, these cells have attracted attention due to the fact they were able to express characteristics of mature intestinal cells, such as enterocytes or mucus producing cells.

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“…Although LS180 cells exhibit basal and inducible CYP3A4 activity (Zheng et al, 2012), they are thought not to form tight junctions in culture. In contrast, T84 cells are known to form tight junctions (Dharmsathaphorn et al, 1984); however, expression of CYP3A4 by T84 cells is controversial (Juuti-Uusitalo et al, 2006;Bourgine et al, 2012), and its inducibility has not been reported. It is worth noting that this cell line expresses pregnane X receptor (PXR), which is one of the important nuclear transcription factors mediating CYP3A4 induction.…”

Section: Introductionmentioning

confidence: 99%

Functional Comparison of Human Colonic Carcinoma Cell Lines and Primary Small Intestinal Epithelial Cells for Investigations of Intestinal Drug Permeability and First-Pass Metabolism

Yamaura

Chapron

Wang

et al. 2015

Drug Metabolism and Disposition

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To further the development of a model for simultaneously assessing intestinal absorption and first-pass metabolism in vitro, Caco-2, LS180, T84, and fetal human small intestinal epithelial cells (fSIECs) were cultured on permeable inserts, and the integrity of cell monolayers, CYP3A4 activity, and the inducibility of enzymes and transporters involved in intestinal drug disposition were measured. Caco-2, T84, and fSIECs all formed tight junctions, as assessed by immunofluorescence microscopy for zonula occludens-1, which was well organized into circumscribing strands in T84, Caco-2, and fSIECs but was diffuse in LS180 cells. The transepithelial electrical resistance value for LS180 monolayers was lower than that for Caco-2, T84, and fSIECs. In addition, the apical-to-basolateral permeability of the paracellular marker Lucifer yellow across LS180 monolayers was greater than in fSIECs, T84, and Caco-2 monolayers. The transcellular marker propranolol exhibited similar permeability across all cells. With regard to metabolic capacity, T84 and LS180 cells showed comparable basal midazolam hydroxylation activity and was inducible by rifampin and 1a,25(OH) 2 D 3 in LS180 cells, but only marginally so in T84 cells. The basal CYP3A4 activity of fSIECs and Caco-2 cells was much lower and not inducible. Interestingly, some of the drug transporters expressed in LS180 and Caco-2 cells were induced by either 1a,25(OH) 2 D 3 or rifampin or both, but effects were limited in the other two cell lines. These results suggest that none of the cell lines tested fully replicated the drug disposition properties of the small intestine and that the search for an ideal screening tool must continue.

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Gene Expression Profiling of Systems Involved in the Metabolism and the Disposition of Xenobiotics: Comparison between Human Intestinal Biopsy Samples and Colon Cell Lines

Cited by 66 publications

References 38 publications

Human Small Intestinal Epithelial Cells Differentiated from Adult Intestinal Stem Cells as a Novel System for Predicting Oral Drug Absorption in Humans

Human Small Intestinal Epithelial Cells Differentiated from Adult Intestinal Stem Cells as a Novel System for Predicting Oral Drug Absorption in Humans

HT29 Cell Line

Functional Comparison of Human Colonic Carcinoma Cell Lines and Primary Small Intestinal Epithelial Cells for Investigations of Intestinal Drug Permeability and First-Pass Metabolism

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