Sucrase-Isomaltase Gene Expression Is Inhibited by Mutant Hepatocyte Nuclear Factor (HNF)-1.ALPHA. and Mutant HNF-1.BETA. in Caco-2 Cells

Gu, Ning; Adachi, Tomohiko; Takeda, Jun; Aoki, Norihiko; Tsujimoto, Gozoh; Ishihara, Akihiko; Tsuda, Kinsuke; Yasuda, Koichiro

doi:10.3177/jnsv.52.105

Cited by 11 publications

(4 citation statements)

References 34 publications

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“…Interestingly, in our study, SI was also detected in HIEC, in contrast to the study by Perreault and Jean-François (6). Therefore, although APN and DPPIV are present on the luminal surface of both crypt and villus cells, which cannot be strictly considered as differentiation markers (33), expression of SI confirmed the differentiated character of HIEC (34). This indicated that, compared with the Perreault and Jean-François study (6), human fetal intestinal epithelial cells showed a more differentiated character in our study, although they may still belong to crypt cells.…”

Section: Discussionsupporting

confidence: 48%

Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3

Liu

Zhang

Zhou

et al. 2010

Braz J Med Biol Res

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Epithelium, a highly dynamic system, plays a key role in the homeostasis of the intestine. However, thus far a human intestinal epithelial cell line has not been established in many countries. Fetal tissue was selected to generate viable cell cultures for its sterile condition, effective generation, and differentiated character. The purpose of the present study was to culture human intestinal epithelial cells by a relatively simple method. Thermolysin was added to improve the yield of epithelial cells, while endothelin-3 was added to stimulate their growth. By adding endothelin-3, the achievement ratio (viable cell cultures/total cultures) was enhanced to 60% of a total of 10 cultures (initiated from 8 distinct fetal small intestines), allowing the generation of viable epithelial cell cultures. Western blot, real-time PCR and immunofluorescent staining showed that cytokeratins 8, 18 and mouse intestinal mucosa-1/39 had high expression levels in human intestinal epithelial cells. Differentiated markers such as sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV also showed high expression levels in human intestinal epithelial cells. Differentiated human intestinal epithelial cells, with the expression of surface markers (cytokeratins 8, 18 and mouse intestinal mucosa-1/39) and secretion of cytokines (sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV), may be cultured by the thermolysin and endothelin-3 method and maintained for at least 20 passages. This is relatively simple, requiring no sophisticated techniques or instruments, and may have a number of varied applications.

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Section: Discussionsupporting

confidence: 48%

Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3

Liu

Zhang

Zhou

et al. 2010

Braz J Med Biol Res

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“…Additionally, in vertical sections, β1 integrin stayed in intracellular regions (presumably Golgi apparatus and granules) until Day 5, and β1 integrin was translocalized into the basolateral membrane at Day 7 (Figure 1A). Accordingly, the expression level of ALPI mRNA, a marker of epithelial cell differentiation and apical membrane (22,23), was significantly increased by Day 7 (Figure 1B). These findings demonstrated that T84 cells successfully gained morphological characteristics of differentiated epithelial cells during the 7-day culture.…”

Section: Resultsmentioning

confidence: 99%

MicroRNA-338-3p and microRNA-451 contribute to the formation of basolateral polarity in epithelial cells

Tsuchiya

Oku

Imanaka

et al. 2009

Nucleic Acids Research

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MicroRNAs are small noncoding RNA species, some of which are playing important roles in cell differentiation. However, the level of participations of microRNAs in epithelial cell differentiation is largely unknown. Here, utilizing an epithelial differentiation model with T84 cells, we demonstrate that miR-338-3p and miR-451 contribute to the formation of epithelial basolateral polarity by facilitating translocalization of β1 integrin to the basolateral membrane. Among 250 microRNAs screened in this study, the expression levels of four microRNAs (miR-33a, 210, 338-3p and 451) were significantly elevated in the differentiated stage of T84 cells, when epithelial cell polarity was established. To investigate the involvement of these microRNAs in terms of epithelial cell polarity, we executed loss-of- and gain-of-function analyses of these microRNAs. The blockade of endogenous miR-338-3p or miR-451 via each microRNA-specific antisense oligonucleotides inhibited the translocalization of β1 integrin to the basolateral membrane, whereas inhibition of miR-210 or miR-33a had no effect on it. On the other hand, simultaneous transfection of synthetic miR-338-3p and miR-451 accelerated the translocalization of β1 integrin to the basolateral membrane, although the introduction of individual synthetic microRNAs exhibited no effect. Therefore, we concluded that both miR-338-3p and miR-451 are necessary for the development of epithelial cell polarity.

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“…Hepatocyte nuclear factor‐1α directly binds to two sites (SIF2, SIF3) of the promoter of the SI gene and upregulates SI gene expression 21 . A reduction in SI gene expression and enzyme activity has been observed in Caco‐2 cells in which a dominant‐negative mutant of HNF‐1α was stably expressed 22 …”

Section: Discussionmentioning

confidence: 99%

GLUCOSE REGULATION OF DIPEPTIDYL PEPTIDASE IV GENE EXPRESSION IS MEDIATED BY HEPATOCYTE NUCLEAR FACTOR‐1α IN EPITHELIAL INTESTINAL CELLS

Tsuda

Matsunaga

et al. 2008

Clin Exp Pharma Physio

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1. Dipeptidyl peptidase IV (DPP-IV) is a new drug target in the treatment of Type 2 diabetes. Dipeptidyl peptidase IV enzyme activity is significantly altered in Type 2 diabetic patients with hyperglycaemia, but the underlying molecular mechanisms remain unclear. 2. The first aim of the present study was to clarify whether glucose regulates DPP-IV enzyme activity. To address this, DPP-IV gene expression and enzyme activity were measured in Caco2 cells cultured in the presence of low (2.5 mmol/L) or high (16.7 mmol/L) concentrations of glucose. We observed that high glucose inhibited DPP-IV gene expression and enzyme activity. 3. The second aim of the present study was to investigate whether hepatocyte nuclear factor (HNF)-1alpha contributes to glucose regulation of DPP-IV gene expression. To explore this question, associations between the gene expression of DPP-IV and HNF-1alpha were examined in Caco-2 cells cultured in the presence of low (2.5 mmol/L) or high (16.7 mmol/L) glucose. We found that the pattern of glucose-regulated DPP-IV gene expression is similar to that of HNF-1alpha. Moreover, to elucidate whether glucose regulation of DPP-IV gene expression is affected when HNF-1alpha is inhibited, we produced two stable cell lines in which a dominant-negative mutant HNF-1alphaR271G or basic vectors were stably expressed. We found that glucose regulation of DPP-IV gene expression was compromised in HNF-1alphaR271G cells, but was well maintained in basic vector cells. 4. These results suggest that glucose regulation of DPP-IV gene expression is mediated by HNF-1alpha.

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Sucrase-Isomaltase Gene Expression Is Inhibited by Mutant Hepatocyte Nuclear Factor (HNF)-1.ALPHA. and Mutant HNF-1.BETA. in Caco-2 Cells

Cited by 11 publications

References 34 publications

Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3

Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3

MicroRNA-338-3p and microRNA-451 contribute to the formation of basolateral polarity in epithelial cells

GLUCOSE REGULATION OF DIPEPTIDYL PEPTIDASE IV GENE EXPRESSION IS MEDIATED BY HEPATOCYTE NUCLEAR FACTOR‐1α IN EPITHELIAL INTESTINAL CELLS

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