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Small Molecule Amiloride Modulates Oncogenic RNA Alternative Splicing to Devitalize Human Cancer Cells

Alternative splicing involves differential exon selection of a gene transcript to generate mRNA and protein isoforms with structural and functional diversity. Abnormal alternative splicing has been shown to be associated with malignant phenotypes of cancer cells, such as chemo-resistance and invasiv...

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Autores principales: Chang, Jan-Gowth, Yang, Den-Mei, Chang, Wen-Hsin, Chow, Lu-Ping, Chan, Wen-Ling, Lin, Hui-Hua, Huang, Hsien-Da, Chang, Ya-Sian, Hung, Cheng-Hao, Yang, Wen-Kuang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3111415/
https://www.ncbi.nlm.nih.gov/pubmed/21694768
http://dx.doi.org/10.1371/journal.pone.0018643
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author Chang, Jan-Gowth
Yang, Den-Mei
Chang, Wen-Hsin
Chow, Lu-Ping
Chan, Wen-Ling
Lin, Hui-Hua
Huang, Hsien-Da
Chang, Ya-Sian
Hung, Cheng-Hao
Yang, Wen-Kuang
author_facet Chang, Jan-Gowth
Yang, Den-Mei
Chang, Wen-Hsin
Chow, Lu-Ping
Chan, Wen-Ling
Lin, Hui-Hua
Huang, Hsien-Da
Chang, Ya-Sian
Hung, Cheng-Hao
Yang, Wen-Kuang
author_sort Chang, Jan-Gowth
collection PubMed
description Alternative splicing involves differential exon selection of a gene transcript to generate mRNA and protein isoforms with structural and functional diversity. Abnormal alternative splicing has been shown to be associated with malignant phenotypes of cancer cells, such as chemo-resistance and invasive activity. Screening small molecules and drugs for modulating RNA splicing in human hepatocellular carcinoma cell line Huh-7, we discovered that amiloride, distinct from four pH-affecting amiloride analogues, could “normalize” the splicing of BCL-X, HIPK3 and RON/MISTR1 transcripts. Our proteomic analyses of amiloride-treated cells detected hypo-phosphorylation of splicing factor SF2/ASF, and decreased levels of SRp20 and two un-identified SR proteins. We further observed decreased phosphorylation of AKT, ERK1/2 and PP1, and increased phosphorylation of p38 and JNK, suggesting that amiloride treatment down-regulates kinases and up-regulates phosphatases in the signal pathways known to affect splicing factor protein phosphorylation. These amiloride effects of “normalized” oncogenic RNA splicing and splicing factor hypo-phosphorylation were both abrogated by pre-treatment with a PP1 inhibitor. Global exon array of amiloride-treated Huh-7 cells detected splicing pattern changes involving 584 exons in 551 gene transcripts, many of which encode proteins playing key roles in ion transport, cellular matrix formation, cytoskeleton remodeling, and genome maintenance. Cellular functional analyses revealed subsequent invasion and migration defects, cell cycle disruption, cytokinesis impairment, and lethal DNA degradation in amiloride-treated Huh-7 cells. Other human solid tumor and leukemic cells, but not a few normal cells, showed similar amiloride-altered RNA splicing with devitalized consequence. This study thus provides mechanistic underpinnings for exploiting small molecule modulation of RNA splicing for cancer therapeutics.
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spelling pubmed-31114152011-06-21 Small Molecule Amiloride Modulates Oncogenic RNA Alternative Splicing to Devitalize Human Cancer Cells Chang, Jan-Gowth Yang, Den-Mei Chang, Wen-Hsin Chow, Lu-Ping Chan, Wen-Ling Lin, Hui-Hua Huang, Hsien-Da Chang, Ya-Sian Hung, Cheng-Hao Yang, Wen-Kuang PLoS One Research Article Alternative splicing involves differential exon selection of a gene transcript to generate mRNA and protein isoforms with structural and functional diversity. Abnormal alternative splicing has been shown to be associated with malignant phenotypes of cancer cells, such as chemo-resistance and invasive activity. Screening small molecules and drugs for modulating RNA splicing in human hepatocellular carcinoma cell line Huh-7, we discovered that amiloride, distinct from four pH-affecting amiloride analogues, could “normalize” the splicing of BCL-X, HIPK3 and RON/MISTR1 transcripts. Our proteomic analyses of amiloride-treated cells detected hypo-phosphorylation of splicing factor SF2/ASF, and decreased levels of SRp20 and two un-identified SR proteins. We further observed decreased phosphorylation of AKT, ERK1/2 and PP1, and increased phosphorylation of p38 and JNK, suggesting that amiloride treatment down-regulates kinases and up-regulates phosphatases in the signal pathways known to affect splicing factor protein phosphorylation. These amiloride effects of “normalized” oncogenic RNA splicing and splicing factor hypo-phosphorylation were both abrogated by pre-treatment with a PP1 inhibitor. Global exon array of amiloride-treated Huh-7 cells detected splicing pattern changes involving 584 exons in 551 gene transcripts, many of which encode proteins playing key roles in ion transport, cellular matrix formation, cytoskeleton remodeling, and genome maintenance. Cellular functional analyses revealed subsequent invasion and migration defects, cell cycle disruption, cytokinesis impairment, and lethal DNA degradation in amiloride-treated Huh-7 cells. Other human solid tumor and leukemic cells, but not a few normal cells, showed similar amiloride-altered RNA splicing with devitalized consequence. This study thus provides mechanistic underpinnings for exploiting small molecule modulation of RNA splicing for cancer therapeutics. Public Library of Science 2011-06-09 /pmc/articles/PMC3111415/ /pubmed/21694768 http://dx.doi.org/10.1371/journal.pone.0018643 Text en Chang et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Chang, Jan-Gowth
Yang, Den-Mei
Chang, Wen-Hsin
Chow, Lu-Ping
Chan, Wen-Ling
Lin, Hui-Hua
Huang, Hsien-Da
Chang, Ya-Sian
Hung, Cheng-Hao
Yang, Wen-Kuang
Small Molecule Amiloride Modulates Oncogenic RNA Alternative Splicing to Devitalize Human Cancer Cells
title Small Molecule Amiloride Modulates Oncogenic RNA Alternative Splicing to Devitalize Human Cancer Cells
title_full Small Molecule Amiloride Modulates Oncogenic RNA Alternative Splicing to Devitalize Human Cancer Cells
title_fullStr Small Molecule Amiloride Modulates Oncogenic RNA Alternative Splicing to Devitalize Human Cancer Cells
title_full_unstemmed Small Molecule Amiloride Modulates Oncogenic RNA Alternative Splicing to Devitalize Human Cancer Cells
title_short Small Molecule Amiloride Modulates Oncogenic RNA Alternative Splicing to Devitalize Human Cancer Cells
title_sort small molecule amiloride modulates oncogenic rna alternative splicing to devitalize human cancer cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3111415/
https://www.ncbi.nlm.nih.gov/pubmed/21694768
http://dx.doi.org/10.1371/journal.pone.0018643
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