miR‐122‐5p promotes aggression and epithelial‐mesenchymal transition in triple‐negative breast cancer by suppressing charged multivesicular body protein 3 through mitogen‐activated protein kinase signaling

Triple‐negative breast cancer (TNBC) is highly metastatic and frequently has a poor prognosis. The lack of comprehension of TNBC and gene therapy targets has led to limitedly effective treatment for TNBC. This study was conducted to better understand the molecular mechanism behind TNBC progression, and to find out promising gene therapy targets for TNBC. Herein the influence of miR‐122‐5p's binding charged multivesicular body protein 3 (CHMP3) 3′‐untranslated region (3′‐UTR) on in TNBC cells was investigated. in vitro experiments quantitative real‐time polymerase chain reaction, immunoblot analysis, dual‐luciferase reporter gene assay, cell counting assay, transwell invasion assay, and flow cytometry‐determined cell apoptosis assay were employed. We also used TargetScan Human 7.2 database to find out the target relationship between miR‐122‐5p and CHMP3 3′‐UTR. TImer algorithm was used to provide an overview of the expression of CHMP3 gene across human pan‐cancer, to predict the survival outcome of breast cancer patients, and to predict the correlation between CHMP3 gene expression and epithelial‐mesenchymal transition (EMT) and mitogen‐activated protein kinase (MAPK)‐related gene expression. CHMP3 gene was significantly downregulated across a wide range of human cancers including breast cancer (BRCA). A higher level of CHMP3 gene predicted a better 3‐ and 5‐year survival outcome of patients with BRCA. In our experiments, miR‐122‐5p was significantly upregulated and CHMP3 gene was significantly downregulated in TNBC cells compared with normal cell line. miR‐122‐5p mimics enhanced TNBC cell viability, proliferation, and invasion whereas the upregulation of CHMP3 gene led to an opposite outcome. Forced expression of miR‐122‐5p suppressed cell apoptosis, compelled EMT and MAPK signaling whereas forced expression of CHMP3 did the opposite. We then conclude that miR‐122‐5p promotes aggression and EMT in TNBC by suppressing CHMP3 through MAPK signaling.

[1]  D. Teis,et al.  The role of the endosomal sorting complexes required for transport (ESCRT) in tumorigenesis , 2014, Molecular membrane biology.

[2]  R. Antoniono,et al.  Neuroendocrine-like differentiation of non-small cell lung carcinoma cells: regulation by cAMP and the interaction of mac25/IGFBP-rP1 and 25.1 , 2006, Oncogene.

[3]  Deling Li,et al.  IMP1 regulates UCA1-mediated cell invasion through facilitating UCA1 decay and decreasing the sponge effect of UCA1 for miR-122-5p , 2018, Breast Cancer Research.

[4]  E. Kohn,et al.  The MAPK pathway across different malignancies: A new perspective , 2014, Cancer.

[5]  K. Vattem,et al.  Performing and optimizing Western blots with an emphasis on chemiluminescent detection. , 2009, Methods in enzymology.

[6]  Samira Mohammadi-Yeganeh,et al.  microRNA as a systemic intervention in the specific breast cancer subtypes with C‐MYC impacts; introducing subtype‐based appraisal tool , 2018, Journal of cellular physiology.

[7]  E. Wilson,et al.  Interaction of IGF-binding protein-related protein 1 with a novel protein, neuroendocrine differentiation factor, results in neuroendocrine differentiation of prostate cancer cells. , 2001, The Journal of clinical endocrinology and metabolism.

[8]  Potential value of circulatory microRNA122 gene expression as a prognostic and metastatic prediction marker for breast cancer , 2019, Molecular Biology Reports.

[9]  G. Lukács,et al.  The ESCRT-III subunit hVps24 is required for degradation but not silencing of the epidermal growth factor receptor. , 2006, Molecular biology of the cell.

[10]  E. Olejniczak,et al.  Myeloid cell leukemia-1 is an important apoptotic survival factor in triple-negative breast cancer , 2015, Cell Death and Differentiation.

[11]  H. Beltran,et al.  The Many Faces of Neuroendocrine Differentiation in Prostate Cancer Progression , 2014, Front. Oncol..

[12]  Y. Uen,et al.  Mining of potential microRNAs with clinical correlation - regulation of syndecan-1 expression by miR-122-5p altered mobility of breast cancer cells and possible correlation with liver injury , 2018, Oncotarget.

[13]  Jorge S Reis-Filho,et al.  Triple-negative breast cancer. , 2010, The New England journal of medicine.

[14]  D. Bailey,et al.  Prognostic and predictive biomarkers in neuroendocrine tumours. , 2017, Critical reviews in oncology/hematology.

[15]  W. Weissenhorn,et al.  Charged Multivesicular Body Protein 2B (CHMP2B) of the Endosomal Sorting Complex Required for Transport-III (ESCRT-III) Polymerizes into Helical Structures Deforming the Plasma Membrane* , 2011, The Journal of Biological Chemistry.

[16]  F. Zhang,et al.  Breast cancer-specific TRAIL expression mediated by miRNA response elements of let-7 and miR-122. , 2014, Neoplasma.

[17]  Hong Wang,et al.  MiR-122 Inhibits Cell Proliferation and Tumorigenesis of Breast Cancer by Targeting IGF1R , 2012, PloS one.

[18]  Sercan Ergün,et al.  The association of the expression of miR-122-5p and its target ADAM10 with human breast cancer , 2014, Molecular Biology Reports.

[19]  S. Emr,et al.  Receptor downregulation and multivesicular-body sorting , 2002, Nature Reviews Molecular Cell Biology.

[20]  N. Tanaka,et al.  Endosomal sorting complex required for transport proteins in cancer pathogenesis, vesicular transport, and non‐endosomal functions , 2008, Cancer science.

[21]  Jianping Chen,et al.  MicroRNA-101 inhibits cell progression and increases paclitaxel sensitivity by suppressing MCL-1 expression in human triple-negative breast cancer , 2015, Oncotarget.

[22]  H. Stenmark,et al.  Cellular Functions and Molecular Mechanisms of the ESCRT Membrane-Scission Machinery. , 2017, Trends in biochemical sciences.

[23]  Qiang Yu,et al.  Protein tyrosine phosphatase UBASH3B is overexpressed in triple-negative breast cancer and promotes invasion and metastasis , 2013, Proceedings of the National Academy of Sciences.

[24]  S. Emr,et al.  The Endosomal Sorting Complex ESCRT-II Mediates the Assembly and Architecture of ESCRT-III Helices , 2012, Cell.

[25]  Jean Gruenberg,et al.  The biogenesis of multivesicular endosomes , 2004, Nature Reviews Molecular Cell Biology.

[26]  Robert A. Weinberg,et al.  EMT in cancer , 2018, Nature Reviews Cancer.

[27]  H. Stenmark,et al.  Defective downregulation of receptor tyrosine kinases in cancer , 2004, The EMBO journal.

[28]  V. Hervieu,et al.  Prognostic factors in neuroendocrine carcinoma: biological markers are more useful than histomorphological markers , 2017, Scientific Reports.

[29]  A. Marchese,et al.  The Endosomal Sorting Complex Required for Transport Pathway Mediates Chemokine Receptor CXCR4-promoted Lysosomal Degradation of the Mammalian Target of Rapamycin Antagonist DEPTOR* , 2015, The Journal of Biological Chemistry.