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Epigenetic Silencing of STAT3-Targeted miR-193a, by Constitutive Activation of JAK/STAT Signaling, Leads to Tumor Progression Through Overexpression of YWHAZ in Gastric Cancer

PURPOSE: The purpose of this study was to identify genes that were epigenetically silenced by STAT3 in gastric cancer. METHODS: MBDcap-Seq and expression microarray were performed to identify genes that were epigenetically silenced in AGS gastric cancer cell lines depleted of STAT3. Cell lines and a...

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Autores principales: Wei, Kuo-Liang, Chou, Jian-Liang, Chen, Yin-Chen, Low, Jie-Ting, Lin, Guan-Ling, Liu, Jing-Lan, Chang, Te-Sheng, Chen, Wei-Ming, Hsieh, Yung-Yu, Yan, Pearlly S., Chuang, Yu-Ming, Lin, Jora M. J., Wu, Shu-Fen, Chiang, Ming-Ko, Li, Chin, Wu, Cheng-Shyong, Chan, Michael W. Y.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7951088/
https://www.ncbi.nlm.nih.gov/pubmed/33718136
http://dx.doi.org/10.3389/fonc.2021.575667
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author Wei, Kuo-Liang
Chou, Jian-Liang
Chen, Yin-Chen
Low, Jie-Ting
Lin, Guan-Ling
Liu, Jing-Lan
Chang, Te-Sheng
Chen, Wei-Ming
Hsieh, Yung-Yu
Yan, Pearlly S.
Chuang, Yu-Ming
Lin, Jora M. J.
Wu, Shu-Fen
Chiang, Ming-Ko
Li, Chin
Wu, Cheng-Shyong
Chan, Michael W. Y.
author_facet Wei, Kuo-Liang
Chou, Jian-Liang
Chen, Yin-Chen
Low, Jie-Ting
Lin, Guan-Ling
Liu, Jing-Lan
Chang, Te-Sheng
Chen, Wei-Ming
Hsieh, Yung-Yu
Yan, Pearlly S.
Chuang, Yu-Ming
Lin, Jora M. J.
Wu, Shu-Fen
Chiang, Ming-Ko
Li, Chin
Wu, Cheng-Shyong
Chan, Michael W. Y.
author_sort Wei, Kuo-Liang
collection PubMed
description PURPOSE: The purpose of this study was to identify genes that were epigenetically silenced by STAT3 in gastric cancer. METHODS: MBDcap-Seq and expression microarray were performed to identify genes that were epigenetically silenced in AGS gastric cancer cell lines depleted of STAT3. Cell lines and animal experiments were performed to investigate proliferation and metastasis of miR-193a and YWHAZ in gastric cancer cell lines. Bisulfite pyrosequencing and tissue microarray were performed to investigate the promoter methylation of miR-193a and expression of STAT3, YWHAZ in patients with gastritis (n = 8) and gastric cancer (n = 71). Quantitative methylation-specific PCR was performed to examine miR-193a promoter methylation in cell-free DNA of serum samples in gastric cancer patients (n = 19). RESULTS: As compared with parental cells, depletion of STAT3 resulted in demethylation of a putative STAT3 target, miR-193a, in AGS gastric cancer cells. Although bisulfite pyrosequencing and epigenetic treatment confirmed that miR-193a was epigenetically silenced in gastric cancer cell lines, ChIP-PCR found that it may be indirectly affected by STAT3. Ectopic expression of miR-193a in AGS cells inhibited proliferation and migration of gastric cancer cells. Further expression microarray and bioinformatics analysis identified YWHAZ as one of the target of miR-193a in AGS gastric cancer cells, such that depletion of YWHAZ reduced migration in AGS cells, while its overexpression increased invasion in MKN45 cells in vitro and in vivo. Clinically, bisulfite pyrosequencing revealed that promoter methylation of miR-193a was significantly higher in human gastric cancer tissues (n = 11) as compared to gastritis (n = 8, p < 0.05). Patients infected with H. pylori showed a significantly higher miR-193a methylation than those without H. pylori infection (p < 0.05). Tissue microarray also showed a positive trend between STAT3 and YWHAZ expression in gastric cancer patients (n = 60). Patients with serum miR-193a methylation was associated with shorter overall survival than those without methylation (p < 0.05). CONCLUSIONS: Constitutive activation of JAK/STAT signaling may confer epigenetic silencing of the STAT3 indirect target and tumor suppressor microRNA, miR-193a in gastric cancer. Transcriptional suppression of miR-193a may led to overexpression of YWHAZ resulting in tumor progression. Targeted inhibition of STAT3 may be a novel therapeutic strategy against gastric cancer.
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spelling pubmed-79510882021-03-12 Epigenetic Silencing of STAT3-Targeted miR-193a, by Constitutive Activation of JAK/STAT Signaling, Leads to Tumor Progression Through Overexpression of YWHAZ in Gastric Cancer Wei, Kuo-Liang Chou, Jian-Liang Chen, Yin-Chen Low, Jie-Ting Lin, Guan-Ling Liu, Jing-Lan Chang, Te-Sheng Chen, Wei-Ming Hsieh, Yung-Yu Yan, Pearlly S. Chuang, Yu-Ming Lin, Jora M. J. Wu, Shu-Fen Chiang, Ming-Ko Li, Chin Wu, Cheng-Shyong Chan, Michael W. Y. Front Oncol Oncology PURPOSE: The purpose of this study was to identify genes that were epigenetically silenced by STAT3 in gastric cancer. METHODS: MBDcap-Seq and expression microarray were performed to identify genes that were epigenetically silenced in AGS gastric cancer cell lines depleted of STAT3. Cell lines and animal experiments were performed to investigate proliferation and metastasis of miR-193a and YWHAZ in gastric cancer cell lines. Bisulfite pyrosequencing and tissue microarray were performed to investigate the promoter methylation of miR-193a and expression of STAT3, YWHAZ in patients with gastritis (n = 8) and gastric cancer (n = 71). Quantitative methylation-specific PCR was performed to examine miR-193a promoter methylation in cell-free DNA of serum samples in gastric cancer patients (n = 19). RESULTS: As compared with parental cells, depletion of STAT3 resulted in demethylation of a putative STAT3 target, miR-193a, in AGS gastric cancer cells. Although bisulfite pyrosequencing and epigenetic treatment confirmed that miR-193a was epigenetically silenced in gastric cancer cell lines, ChIP-PCR found that it may be indirectly affected by STAT3. Ectopic expression of miR-193a in AGS cells inhibited proliferation and migration of gastric cancer cells. Further expression microarray and bioinformatics analysis identified YWHAZ as one of the target of miR-193a in AGS gastric cancer cells, such that depletion of YWHAZ reduced migration in AGS cells, while its overexpression increased invasion in MKN45 cells in vitro and in vivo. Clinically, bisulfite pyrosequencing revealed that promoter methylation of miR-193a was significantly higher in human gastric cancer tissues (n = 11) as compared to gastritis (n = 8, p < 0.05). Patients infected with H. pylori showed a significantly higher miR-193a methylation than those without H. pylori infection (p < 0.05). Tissue microarray also showed a positive trend between STAT3 and YWHAZ expression in gastric cancer patients (n = 60). Patients with serum miR-193a methylation was associated with shorter overall survival than those without methylation (p < 0.05). CONCLUSIONS: Constitutive activation of JAK/STAT signaling may confer epigenetic silencing of the STAT3 indirect target and tumor suppressor microRNA, miR-193a in gastric cancer. Transcriptional suppression of miR-193a may led to overexpression of YWHAZ resulting in tumor progression. Targeted inhibition of STAT3 may be a novel therapeutic strategy against gastric cancer. Frontiers Media S.A. 2021-02-25 /pmc/articles/PMC7951088/ /pubmed/33718136 http://dx.doi.org/10.3389/fonc.2021.575667 Text en Copyright © 2021 Wei, Chou, Chen, Low, Lin, Liu, Chang, Chen, Hsieh, Yan, Chuang, Lin, Wu, Chiang, Li, Wu and Chan http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Oncology
Wei, Kuo-Liang
Chou, Jian-Liang
Chen, Yin-Chen
Low, Jie-Ting
Lin, Guan-Ling
Liu, Jing-Lan
Chang, Te-Sheng
Chen, Wei-Ming
Hsieh, Yung-Yu
Yan, Pearlly S.
Chuang, Yu-Ming
Lin, Jora M. J.
Wu, Shu-Fen
Chiang, Ming-Ko
Li, Chin
Wu, Cheng-Shyong
Chan, Michael W. Y.
Epigenetic Silencing of STAT3-Targeted miR-193a, by Constitutive Activation of JAK/STAT Signaling, Leads to Tumor Progression Through Overexpression of YWHAZ in Gastric Cancer
title Epigenetic Silencing of STAT3-Targeted miR-193a, by Constitutive Activation of JAK/STAT Signaling, Leads to Tumor Progression Through Overexpression of YWHAZ in Gastric Cancer
title_full Epigenetic Silencing of STAT3-Targeted miR-193a, by Constitutive Activation of JAK/STAT Signaling, Leads to Tumor Progression Through Overexpression of YWHAZ in Gastric Cancer
title_fullStr Epigenetic Silencing of STAT3-Targeted miR-193a, by Constitutive Activation of JAK/STAT Signaling, Leads to Tumor Progression Through Overexpression of YWHAZ in Gastric Cancer
title_full_unstemmed Epigenetic Silencing of STAT3-Targeted miR-193a, by Constitutive Activation of JAK/STAT Signaling, Leads to Tumor Progression Through Overexpression of YWHAZ in Gastric Cancer
title_short Epigenetic Silencing of STAT3-Targeted miR-193a, by Constitutive Activation of JAK/STAT Signaling, Leads to Tumor Progression Through Overexpression of YWHAZ in Gastric Cancer
title_sort epigenetic silencing of stat3-targeted mir-193a, by constitutive activation of jak/stat signaling, leads to tumor progression through overexpression of ywhaz in gastric cancer
topic Oncology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7951088/
https://www.ncbi.nlm.nih.gov/pubmed/33718136
http://dx.doi.org/10.3389/fonc.2021.575667
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