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The miR-203a Regulatory Network Affects the Proliferation of Chronic Myeloid Leukemia K562 Cells

To study the molecular mechanism by which miR-203a affects the development of CML, bioinformatics software was used to predict the upstream transcription factors and downstream target genes of miR-203a. A 5’-rapid amplification of cDNA ends assay was performed to detect gene transcription initiation...

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Autores principales: He, Jinhua, Han, Zeping, An, Ziyi, Li, Yumin, Xie, Xingyi, Zhou, Jiabin, He, Sihua, Lv, Yubing, He, Mengling, Qu, Hong, Liu, Gexiu, Li, Yuguang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7917221/
https://www.ncbi.nlm.nih.gov/pubmed/33659248
http://dx.doi.org/10.3389/fcell.2021.616711
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author He, Jinhua
Han, Zeping
An, Ziyi
Li, Yumin
Xie, Xingyi
Zhou, Jiabin
He, Sihua
Lv, Yubing
He, Mengling
Qu, Hong
Liu, Gexiu
Li, Yuguang
author_facet He, Jinhua
Han, Zeping
An, Ziyi
Li, Yumin
Xie, Xingyi
Zhou, Jiabin
He, Sihua
Lv, Yubing
He, Mengling
Qu, Hong
Liu, Gexiu
Li, Yuguang
author_sort He, Jinhua
collection PubMed
description To study the molecular mechanism by which miR-203a affects the development of CML, bioinformatics software was used to predict the upstream transcription factors and downstream target genes of miR-203a. A 5’-rapid amplification of cDNA ends assay was performed to detect gene transcription initiation sites. A chromatin immunoprecipitation assay was used to verify the binding of transcription factors and promoter regions. A double luciferase reporter gene vector was constructed to demonstrate the regulatory effect of miR-203a on target genes. Real-time PCR and western blotting were used to detect the relative expression levels of genes and proteins, respectively. The results showed that there was a binding site for the transcription factor EGR1 in the upstream promoter region of miR-203a. WT1, BMI1, and XIAP were identified as target genes regulated by miR-203a. EGR1 and miR-203a were downregulated in human peripheral blood mononuclear cells and the CML K562 cell line, while WT1, BMI1, and XIAP were upregulated. The transcription initiation site of miR-203a was identified in the upstream promoter region (G nucleotide at −339 bp), and the transcription factor EGR1 could bind to the promoter region (at −268 bp) of miR-203a and increase its expression. Over expression of miR-203a inhibited the proliferation of K562 cells. A rescue assay showed that overexpression of WT1, BMI1, and XIAP offset the antitumor effect of miR-203a. Conclusion, EGR1 positively regulated the expression of miR-203a, thus relieving the inhibition of miR-203a on the translation of its target genes (WT1, BMI1, and XIAP) and affecting the proliferation of K562 cells.
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spelling pubmed-79172212021-03-02 The miR-203a Regulatory Network Affects the Proliferation of Chronic Myeloid Leukemia K562 Cells He, Jinhua Han, Zeping An, Ziyi Li, Yumin Xie, Xingyi Zhou, Jiabin He, Sihua Lv, Yubing He, Mengling Qu, Hong Liu, Gexiu Li, Yuguang Front Cell Dev Biol Cell and Developmental Biology To study the molecular mechanism by which miR-203a affects the development of CML, bioinformatics software was used to predict the upstream transcription factors and downstream target genes of miR-203a. A 5’-rapid amplification of cDNA ends assay was performed to detect gene transcription initiation sites. A chromatin immunoprecipitation assay was used to verify the binding of transcription factors and promoter regions. A double luciferase reporter gene vector was constructed to demonstrate the regulatory effect of miR-203a on target genes. Real-time PCR and western blotting were used to detect the relative expression levels of genes and proteins, respectively. The results showed that there was a binding site for the transcription factor EGR1 in the upstream promoter region of miR-203a. WT1, BMI1, and XIAP were identified as target genes regulated by miR-203a. EGR1 and miR-203a were downregulated in human peripheral blood mononuclear cells and the CML K562 cell line, while WT1, BMI1, and XIAP were upregulated. The transcription initiation site of miR-203a was identified in the upstream promoter region (G nucleotide at −339 bp), and the transcription factor EGR1 could bind to the promoter region (at −268 bp) of miR-203a and increase its expression. Over expression of miR-203a inhibited the proliferation of K562 cells. A rescue assay showed that overexpression of WT1, BMI1, and XIAP offset the antitumor effect of miR-203a. Conclusion, EGR1 positively regulated the expression of miR-203a, thus relieving the inhibition of miR-203a on the translation of its target genes (WT1, BMI1, and XIAP) and affecting the proliferation of K562 cells. Frontiers Media S.A. 2021-02-15 /pmc/articles/PMC7917221/ /pubmed/33659248 http://dx.doi.org/10.3389/fcell.2021.616711 Text en Copyright © 2021 He, Han, An, Li, Xie, Zhou, He, Lv, He, Qu, Liu and Li. 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 Cell and Developmental Biology
He, Jinhua
Han, Zeping
An, Ziyi
Li, Yumin
Xie, Xingyi
Zhou, Jiabin
He, Sihua
Lv, Yubing
He, Mengling
Qu, Hong
Liu, Gexiu
Li, Yuguang
The miR-203a Regulatory Network Affects the Proliferation of Chronic Myeloid Leukemia K562 Cells
title The miR-203a Regulatory Network Affects the Proliferation of Chronic Myeloid Leukemia K562 Cells
title_full The miR-203a Regulatory Network Affects the Proliferation of Chronic Myeloid Leukemia K562 Cells
title_fullStr The miR-203a Regulatory Network Affects the Proliferation of Chronic Myeloid Leukemia K562 Cells
title_full_unstemmed The miR-203a Regulatory Network Affects the Proliferation of Chronic Myeloid Leukemia K562 Cells
title_short The miR-203a Regulatory Network Affects the Proliferation of Chronic Myeloid Leukemia K562 Cells
title_sort mir-203a regulatory network affects the proliferation of chronic myeloid leukemia k562 cells
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7917221/
https://www.ncbi.nlm.nih.gov/pubmed/33659248
http://dx.doi.org/10.3389/fcell.2021.616711
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