MicroRNA-101 (miR-101) post-transcriptionally regulates the expression of EP4 receptor in colon cancers

Anupama Chandramouli, Benjamin Chidi Onyeagucha, Melania E. Mercado-Pimentel, Lenka Stankova, Nisreen Abu Shahin, Bonnie J. LaFleur, Ronald L. Heimark, Achyut K. Bhattacharyya, Mark A. Nelson

Research output: Contribution to journalArticle

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Abstract

Purpose: Expression of the PGE2 receptor, EP4, is up-regulated during colorectal carcinogenesis. However the mechanism leading to deregulation of the EP4 receptor is not known. The present study was conducted to investigate the regulation of EP4 receptor by miRNAs. Experimental Design: We analyzed 26 colon cancers (i.e. 15 adenocarcinomas and 9 adenomas) and 16 normal colon specimens for EP4 receptor expression by immunohistochemistry. A bioinformatics approached identified putative microRNA binding sites with the 3′-UTR of the EP4 receptor. Both colon cancer cell lines and tumor specimens were analyzed for miR-101 and EP4 expression by qRT-PCR and Western analysis respectively and simultaneously in situ hybridizations was used to confirm our results. In vitro and in vivo assays were used to confirm our clinical findings. Results: We observed an inverse correlation between the levels of miR-101 and EP4 receptor protein. Transfection of LS174T cells with miR-101 significantly suppressed a luciferase reporter containing the EP4 receptor-3′-UTR. In contrast, a mutant EP4 receptor-3′-UTR construct was unaffected. Ectopic expression of miR-101 markedly reduced cell proliferation and motility. Co-transfection of EP4 receptor could rescue colon cancer cells from the tumor suppressive effects of miR-101. Moreover, the pharmacologic inhibition of EP4 receptor signaling or silencing of EP4 receptor phenocopied the effect of miR-101. This is the first study to show that the EP4 receptor is negatively regulated by miR-101. Conclusions: These data provide new insights in the modulation of EP-4 receptor expression at the post-transcriptional level by miR-101 and suggests therapeutic strategies against miR-101 targets may be warranted.

Original languageEnglish (US)
Pages (from-to)175-184
Number of pages10
JournalCancer Biology and Therapy
Volume13
Issue number3
DOIs
StatePublished - Feb 1 2012

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MicroRNAs
Colonic Neoplasms
3' Untranslated Regions
Transfection
Prostaglandin E Receptors
Computational Biology
Tumor Cell Line
Luciferases
Adenoma
Cell Movement
In Situ Hybridization
Colon
Carcinogenesis
Adenocarcinoma
Research Design
Immunohistochemistry
Binding Sites
Cell Proliferation
Polymerase Chain Reaction

Keywords

  • Colorectal cancer
  • EP4
  • miR-101
  • Post-transcriptional regulation

ASJC Scopus subject areas

  • Cancer Research
  • Oncology
  • Molecular Medicine
  • Pharmacology

Cite this

Chandramouli, A., Onyeagucha, B. C., Mercado-Pimentel, M. E., Stankova, L., Abu Shahin, N., LaFleur, B. J., ... Nelson, M. A. (2012). MicroRNA-101 (miR-101) post-transcriptionally regulates the expression of EP4 receptor in colon cancers. Cancer Biology and Therapy, 13(3), 175-184. https://doi.org/10.4161/cbt.13.3.18874

MicroRNA-101 (miR-101) post-transcriptionally regulates the expression of EP4 receptor in colon cancers. / Chandramouli, Anupama; Onyeagucha, Benjamin Chidi; Mercado-Pimentel, Melania E.; Stankova, Lenka; Abu Shahin, Nisreen; LaFleur, Bonnie J.; Heimark, Ronald L.; Bhattacharyya, Achyut K.; Nelson, Mark A.

In: Cancer Biology and Therapy, Vol. 13, No. 3, 01.02.2012, p. 175-184.

Research output: Contribution to journalArticle

Chandramouli, A, Onyeagucha, BC, Mercado-Pimentel, ME, Stankova, L, Abu Shahin, N, LaFleur, BJ, Heimark, RL, Bhattacharyya, AK & Nelson, MA 2012, 'MicroRNA-101 (miR-101) post-transcriptionally regulates the expression of EP4 receptor in colon cancers', Cancer Biology and Therapy, vol. 13, no. 3, pp. 175-184. https://doi.org/10.4161/cbt.13.3.18874
Chandramouli, Anupama ; Onyeagucha, Benjamin Chidi ; Mercado-Pimentel, Melania E. ; Stankova, Lenka ; Abu Shahin, Nisreen ; LaFleur, Bonnie J. ; Heimark, Ronald L. ; Bhattacharyya, Achyut K. ; Nelson, Mark A. / MicroRNA-101 (miR-101) post-transcriptionally regulates the expression of EP4 receptor in colon cancers. In: Cancer Biology and Therapy. 2012 ; Vol. 13, No. 3. pp. 175-184.
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abstract = "Purpose: Expression of the PGE2 receptor, EP4, is up-regulated during colorectal carcinogenesis. However the mechanism leading to deregulation of the EP4 receptor is not known. The present study was conducted to investigate the regulation of EP4 receptor by miRNAs. Experimental Design: We analyzed 26 colon cancers (i.e. 15 adenocarcinomas and 9 adenomas) and 16 normal colon specimens for EP4 receptor expression by immunohistochemistry. A bioinformatics approached identified putative microRNA binding sites with the 3′-UTR of the EP4 receptor. Both colon cancer cell lines and tumor specimens were analyzed for miR-101 and EP4 expression by qRT-PCR and Western analysis respectively and simultaneously in situ hybridizations was used to confirm our results. In vitro and in vivo assays were used to confirm our clinical findings. Results: We observed an inverse correlation between the levels of miR-101 and EP4 receptor protein. Transfection of LS174T cells with miR-101 significantly suppressed a luciferase reporter containing the EP4 receptor-3′-UTR. In contrast, a mutant EP4 receptor-3′-UTR construct was unaffected. Ectopic expression of miR-101 markedly reduced cell proliferation and motility. Co-transfection of EP4 receptor could rescue colon cancer cells from the tumor suppressive effects of miR-101. Moreover, the pharmacologic inhibition of EP4 receptor signaling or silencing of EP4 receptor phenocopied the effect of miR-101. This is the first study to show that the EP4 receptor is negatively regulated by miR-101. Conclusions: These data provide new insights in the modulation of EP-4 receptor expression at the post-transcriptional level by miR-101 and suggests therapeutic strategies against miR-101 targets may be warranted.",
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AU - Onyeagucha, Benjamin Chidi

AU - Mercado-Pimentel, Melania E.

AU - Stankova, Lenka

AU - Abu Shahin, Nisreen

AU - LaFleur, Bonnie J.

AU - Heimark, Ronald L.

AU - Bhattacharyya, Achyut K.

AU - Nelson, Mark A.

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N2 - Purpose: Expression of the PGE2 receptor, EP4, is up-regulated during colorectal carcinogenesis. However the mechanism leading to deregulation of the EP4 receptor is not known. The present study was conducted to investigate the regulation of EP4 receptor by miRNAs. Experimental Design: We analyzed 26 colon cancers (i.e. 15 adenocarcinomas and 9 adenomas) and 16 normal colon specimens for EP4 receptor expression by immunohistochemistry. A bioinformatics approached identified putative microRNA binding sites with the 3′-UTR of the EP4 receptor. Both colon cancer cell lines and tumor specimens were analyzed for miR-101 and EP4 expression by qRT-PCR and Western analysis respectively and simultaneously in situ hybridizations was used to confirm our results. In vitro and in vivo assays were used to confirm our clinical findings. Results: We observed an inverse correlation between the levels of miR-101 and EP4 receptor protein. Transfection of LS174T cells with miR-101 significantly suppressed a luciferase reporter containing the EP4 receptor-3′-UTR. In contrast, a mutant EP4 receptor-3′-UTR construct was unaffected. Ectopic expression of miR-101 markedly reduced cell proliferation and motility. Co-transfection of EP4 receptor could rescue colon cancer cells from the tumor suppressive effects of miR-101. Moreover, the pharmacologic inhibition of EP4 receptor signaling or silencing of EP4 receptor phenocopied the effect of miR-101. This is the first study to show that the EP4 receptor is negatively regulated by miR-101. Conclusions: These data provide new insights in the modulation of EP-4 receptor expression at the post-transcriptional level by miR-101 and suggests therapeutic strategies against miR-101 targets may be warranted.

AB - Purpose: Expression of the PGE2 receptor, EP4, is up-regulated during colorectal carcinogenesis. However the mechanism leading to deregulation of the EP4 receptor is not known. The present study was conducted to investigate the regulation of EP4 receptor by miRNAs. Experimental Design: We analyzed 26 colon cancers (i.e. 15 adenocarcinomas and 9 adenomas) and 16 normal colon specimens for EP4 receptor expression by immunohistochemistry. A bioinformatics approached identified putative microRNA binding sites with the 3′-UTR of the EP4 receptor. Both colon cancer cell lines and tumor specimens were analyzed for miR-101 and EP4 expression by qRT-PCR and Western analysis respectively and simultaneously in situ hybridizations was used to confirm our results. In vitro and in vivo assays were used to confirm our clinical findings. Results: We observed an inverse correlation between the levels of miR-101 and EP4 receptor protein. Transfection of LS174T cells with miR-101 significantly suppressed a luciferase reporter containing the EP4 receptor-3′-UTR. In contrast, a mutant EP4 receptor-3′-UTR construct was unaffected. Ectopic expression of miR-101 markedly reduced cell proliferation and motility. Co-transfection of EP4 receptor could rescue colon cancer cells from the tumor suppressive effects of miR-101. Moreover, the pharmacologic inhibition of EP4 receptor signaling or silencing of EP4 receptor phenocopied the effect of miR-101. This is the first study to show that the EP4 receptor is negatively regulated by miR-101. Conclusions: These data provide new insights in the modulation of EP-4 receptor expression at the post-transcriptional level by miR-101 and suggests therapeutic strategies against miR-101 targets may be warranted.

KW - Colorectal cancer

KW - EP4

KW - miR-101

KW - Post-transcriptional regulation

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