The miR-203a Regulatory Network Affects the Proliferation of Chronic Myeloid Leukemia K562 Cells
Open Access
- 15 February 2021
- journal article
- research article
- Published by Frontiers Media SA in Frontiers in Cell and Developmental Biology
Abstract
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.Funding Information
- Natural Science Foundation of Guangdong Province
This publication has 32 references indexed in Scilit:
- Imatinib induces demethylation of miR-203 gene: An epigenetic mechanism of anti-tumor effect of imatinibLeukemia Research, 2013
- hsa-miR-203 enhances the sensitivity of leukemia cells to arsenic trioxideExperimental and Therapeutic Medicine, 2013
- Consistency Test of the Cell Cycle: Roles for p53 and EGR1Cancer Research, 2012
- Epigenetic inactivation of the hsa-miR-203 in haematological malignanciesJournal of Cellular and Molecular Medicine, 2011
- BMI-1 Autoantibody as a New Potential Biomarker for Cervical CarcinomaPLOS ONE, 2011
- Simultaneous targeting of P‐gp and XIAP with siRNAs increases sensitivity of P‐gp overexpressing CML cells to imatinibHematology, 2011
- Micro-RNA response to imatinib mesylate in patients with chronic myeloid leukemiaHaematologica, 2010
- Genetic and Epigenetic Silencing of MicroRNA-203 Enhances ABL1 and BCR-ABL1 Oncogene ExpressionCancer Cell, 2008
- The role of the Wilms tumour gene (WT1) in normal and malignant haematopoiesisExpert Reviews in Molecular Medicine, 2007
- Chemosensitization Effects of XIAP Downregulation in K562 Leukemia CellsJournal of Chemotherapy, 2006