Downregulation of NHE-3 (SLC9A3) expression by MicroRNAs in intestinal epithelial cells
Na+/H+ exchanger-3 (NHE-3) is the major apical membrane transporter involved in vectorial Na+ absorption in the intestine. Dysregulation of NHE-3 expression and/or function has been implicated in pathophysiology of diarrhea associated with gut inflammation and infections. Therefore, it is critical to understand the mechanisms involved in the regulation of NHE-3 expression. MicroRNAs (miRNAs) are highly conserved small RNAs that can regulate gene expression at the posttranscriptional level. To date, however, very little is known about the regulation of NHE-3 expression by microRNAs. Therefore, current studies were undertaken to examine the potential miRNA candidates that can regulate the expression of NHE-3 in intestinal epithelial cells. In silico analysis, using different algorithms, predicted several miRNAs that target NHE-3. MicroRNAs with highest context and target score, miR-326, miR-744-5p, and miR-330-5p, were selected for the current study. Human NHE-3 gene 3' untranslated region [3'UTR; 160 base pair (bp)] was cloned into pmirGLO vector upstream of luciferase reporter and transiently transfected with mimics of miR-326, miR-744-5p, and miR-330-5p into Caco-2, HT-29, and SK-CO15 cells. Cotransfection of NHE-3 3' UTR with miR-326 and -miR-330-5p mimics resulted in a significant decrease in relative luciferase activity. Transfection of miR-326 and -330-5p mimics into SK-CO15 cells significantly decreased the NHE-3 protein expression, with no change in NHE-3 messenger ribonucleic acid (mRNA) levels. Our findings demonstrate a novel mechanism for posttranscriptional regulation of NHE-3 by miR-326 and -330-5p by translational repression. We speculate that miR-326 and -330-5p dependent pathways may be involved in modulating NHE-3 expression under physiological and pathophysiological conditions.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:323 |
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| Enthalten in: |
American journal of physiology. Cell physiology - 323(2022), 6 vom: 01. Dez., Seite C1720-C1727 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Anbazhagan, Arivarasu N [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 09.12.2022 Date Revised 02.12.2023 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1152/ajpcell.00294.2022 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM347054609 |
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| 100 | 1 | |a Anbazhagan, Arivarasu N |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Downregulation of NHE-3 (SLC9A3) expression by MicroRNAs in intestinal epithelial cells |
| 264 | 1 | |c 2022 | |
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| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Na+/H+ exchanger-3 (NHE-3) is the major apical membrane transporter involved in vectorial Na+ absorption in the intestine. Dysregulation of NHE-3 expression and/or function has been implicated in pathophysiology of diarrhea associated with gut inflammation and infections. Therefore, it is critical to understand the mechanisms involved in the regulation of NHE-3 expression. MicroRNAs (miRNAs) are highly conserved small RNAs that can regulate gene expression at the posttranscriptional level. To date, however, very little is known about the regulation of NHE-3 expression by microRNAs. Therefore, current studies were undertaken to examine the potential miRNA candidates that can regulate the expression of NHE-3 in intestinal epithelial cells. In silico analysis, using different algorithms, predicted several miRNAs that target NHE-3. MicroRNAs with highest context and target score, miR-326, miR-744-5p, and miR-330-5p, were selected for the current study. Human NHE-3 gene 3' untranslated region [3'UTR; 160 base pair (bp)] was cloned into pmirGLO vector upstream of luciferase reporter and transiently transfected with mimics of miR-326, miR-744-5p, and miR-330-5p into Caco-2, HT-29, and SK-CO15 cells. Cotransfection of NHE-3 3' UTR with miR-326 and -miR-330-5p mimics resulted in a significant decrease in relative luciferase activity. Transfection of miR-326 and -330-5p mimics into SK-CO15 cells significantly decreased the NHE-3 protein expression, with no change in NHE-3 messenger ribonucleic acid (mRNA) levels. Our findings demonstrate a novel mechanism for posttranscriptional regulation of NHE-3 by miR-326 and -330-5p by translational repression. We speculate that miR-326 and -330-5p dependent pathways may be involved in modulating NHE-3 expression under physiological and pathophysiological conditions | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, U.S. Gov't, Non-P.H.S. | |
| 650 | 4 | |a intestinal ion transporters | |
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| 650 | 7 | |a Sodium-Hydrogen Exchanger 3 |2 NLM | |
| 650 | 7 | |a SLC9A3 protein, human |2 NLM | |
| 700 | 1 | |a Priyamvada, Shubha |e verfasserin |4 aut | |
| 700 | 1 | |a Kumar, Anoop |e verfasserin |4 aut | |
| 700 | 1 | |a Jayawardena, Dulari |e verfasserin |4 aut | |
| 700 | 1 | |a Borthakur, Alip |e verfasserin |4 aut | |
| 700 | 1 | |a Gill, Ravinder K |e verfasserin |4 aut | |
| 700 | 1 | |a Alrefai, Waddah A |e verfasserin |4 aut | |
| 700 | 1 | |a Dudeja, Pradeep K |e verfasserin |4 aut | |
| 700 | 1 | |a Saksena, Seema |e verfasserin |4 aut | |
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