MicroRNA Expression Profiling on Paired Primary and Lymph Node Metastatic Breast Cancer Revealed Distinct microRNA Profile Associated With LNM

Breast cancer (BC) is the foremost cause of cancer-related deaths in women. BC patients are oftentimes presented with lymph node metastasis (LNM), which increases their risk of recurrence. Compelling data have recently implicated microRNAs in promoting BC metastasis. Therefore, the identification of microRNA (miRNA)-based molecular signature associated with LNM could provide an opportunity for a more personalized treatment for BC patients with high risk of LNM. In current study, we performed comprehensive miRNA profiling in matched primary breast and LNM and identified 40 miRNAs, which were differentially expressed in LNM compared to primary tumors. The expression of 14 miRNAs (Up: hsa-miR-155-5p, hsa-miR-150-5p, hsa-miR-146a-5p, hsa-miR-142-5p and down: hsa-miR-200a-3p, hsa-miR-200b-3p, hsa-miR-200c-3p, hsa-miR-205-5p, hsa-miR-210-3p, hsa-miR-214-3p, hsa-miR-141-3p, hsa-miR-127-3p, hsa-miR-125a-5p, and hsa-let-7c-5p) was subsequently validated in a second cohort of 32 breast and 32 matched LNM tumor tissues. Mechanistically, forced expression of hsa-miR-205-5p, or hsa-miR-214-3p epigenetically inhibited MDA-MB-231 cell proliferation, colony formation, and cell migration. Global gene expression profiling on MDA-MB-231 cells overexpressing hsa-miR-205-5p, or hsa-miR-214-3p in combination with in silico target prediction and ingenuity pathway analyses identified multiple bona fide targets for hsa-miR-205-5p, hsa-miR-214-3p affecting cellular proliferation and migration. Interestingly, interrogation of the expression levels of hsa-miR-205 and hsa-miR-214 in the METABRIC breast cancer dataset revealed significantly poor overall survival in patients with downregulated expression of miR-205 [HR = 0.75 (0.61–0.91)], p = 0.003 and hsa-miR-214 [HR = 0.74 (0.59–0.93) p = 0.008]. Our data unraveled the miRNA-transcriptional landscape associated with LNM and provide novel insight on the role of several miRNAs in promoting BC LNM, and suggest their potential utilization in the clinical management of BC patients.

[1]  Paula D. Bos,et al.  Metastasis: from dissemination to organ-specific colonization , 2009, Nature Reviews Cancer.

[2]  Hideaki Sugawara,et al.  The Sequence Read Archive , 2010, Nucleic Acids Res..

[3]  E. Nikitina,et al.  MicroRNAs and human cancer. , 2012, Experimental oncology.

[4]  N. Alajez,et al.  Noncoding RNAs as potential mediators of resistance to cancer immunotherapy. , 2019, Seminars in cancer biology.

[5]  S. Rana,et al.  MicroRNAs in Cancer: Challenges and Opportunities in Early Detection, Disease Monitoring, and Therapeutic Agents , 2017, Current Pathobiology Reports.

[6]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[7]  M. Kassem,et al.  Neoplastic Transformation of Human Mesenchymal Stromal Cells Mediated via LIN28B , 2019, Scientific Reports.

[8]  Stefan Wiemann,et al.  MicroRNA-200c Represses Migration and Invasion of Breast Cancer Cells by Targeting Actin-Regulatory Proteins FHOD1 and PPM1F , 2011, Molecular and Cellular Biology.

[9]  J. Peterse,et al.  Breast cancer metastasis: markers and models , 2005, Nature Reviews Cancer.

[10]  F. de Longueville,et al.  Comparative clinical and transcriptomal profiles of breast cancer between French and South Mediterranean patients show minor but significative biological differences. , 2008, Cancer genomics & proteomics.

[11]  Lin He,et al.  MicroRNAs: small RNAs with a big role in gene regulation , 2004, Nature reviews genetics.

[12]  George P Cobb,et al.  microRNAs as oncogenes and tumor suppressors. , 2007, Developmental biology.

[13]  I. Berindan‐Neagoe,et al.  The silent healer: miR-205-5p up-regulation inhibits epithelial to mesenchymal transition in colon cancer cells by indirectly up-regulating E-cadherin expression , 2018, Cell Death & Disease.

[14]  Larry Norton,et al.  Tumor Self-Seeding by Circulating Cancer Cells , 2009, Cell.

[15]  Yuepu Pu,et al.  The Cluster of miR-143 and miR-145 Affects the Risk for Esophageal Squamous Cell Carcinoma through Co-Regulating Fascin Homolog 1 , 2012, PloS one.

[16]  M. Esteller.  Non-coding RNAs in human disease , 2011, Nature Reviews Genetics.

[17]  M. Seoud,et al.  Trends in epidemiology and management of breast cancer in developing Arab countries: a literature and registry analysis. , 2007, International journal of surgery.

[18]  P. Kronqvist,et al.  Proliferation-associated miRNAs-494, -205, -21 and -126 detected by in situ hybridization: expression and prognostic potential in breast carcinoma patients , 2018, Journal of Cancer Research and Clinical Oncology.

[19]  M. Kassem,et al.  Genome-wide mRNA and miRNA expression profiling reveal multiple regulatory networks in colorectal cancer , 2015, Cell Death and Disease.

[20]  Wenjie Zhu,et al.  A 4-microRNA signature predicts lymph node metastasis and prognosis in breast cancer. , 2018, Human pathology.

[21]  A. Jemal,et al.  Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries , 2018, CA: a cancer journal for clinicians.

[22]  Isabella Castiglioni,et al.  MicroRNAs: New Biomarkers for Diagnosis, Prognosis, Therapy Prediction and Therapeutic Tools for Breast Cancer , 2015, Theranostics.

[23]  A. Hui,et al.  MiR-218 suppresses nasopharyngeal cancer progression through downregulation of survivin and the SLIT2-ROBO1 pathway. , 2011, Cancer research.

[24]  Emad A. Ahmed,et al.  SOX4: Epigenetic Regulation and Role in Tumorigenesis. , 2020, Seminars in cancer biology.

[25]  M. Korpal,et al.  The miR-200 Family Inhibits Epithelial-Mesenchymal Transition and Cancer Cell Migration by Direct Targeting of E-cadherin Transcriptional Repressors ZEB1 and ZEB2* , 2008, Journal of Biological Chemistry.

[26]  Moustapha Kassem,et al.  microRNA expression profiling on individual breast cancer patients identifies novel panel of circulating microRNA for early detection , 2016, Scientific Reports.

[27]  M. Kassem,et al.  Pleiotropic effects of cancer cells’ secreted factors on human stromal (mesenchymal) stem cells , 2013, Stem Cell Research & Therapy.

[28]  M. Ashrafizadeh,et al.  MicroRNAs as novel targets of sulforaphane in cancer therapy: The beginning of a new tale? , 2020, Phytotherapy research : PTR.

[29]  Erratum: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. , 2020, CA: a cancer journal for clinicians.

[30]  P. Lønning,et al.  Genomic Evolution of Breast Cancer Metastasis and Relapse , 2017, Cancer cell.

[31]  F. Markowetz,et al.  The genomic and transcriptomic architecture of 2,000 breast tumours reveals novel subgroups , 2012, Nature.

[32]  Ignace Vergote,et al.  Dysregulation of microRNAs in breast cancer and their potential role as prognostic and predictive biomarkers in patient management , 2015, Breast Cancer Research.

[33]  N. Alajez,et al.  Integrated Transcriptome and Pathway Analyses Revealed Multiple Activated Pathways in Breast Cancer , 2019, Front. Oncol..

[34]  E. Butcher,et al.  Chemokines in tissue-specific and microenvironment-specific lymphocyte homing. , 2000, Current opinion in immunology.

[35]  Michael F. Clarke,et al.  Downregulation of miRNA-200c Links Breast Cancer Stem Cells with Normal Stem Cells , 2009, Cell.

[36]  R. Ouellette,et al.  MiRNAs 484 and 210 Regulate Pax-5 Expression and Function in Breast Cancer Cells. , 2019, Carcinogenesis.

[37]  K. Pantel,et al.  Cancer micrometastases , 2009, Nature Reviews Clinical Oncology.

[38]  Lotfi Chouchane,et al.  Breast cancer in Arab populations: molecular characteristics and disease management implications. , 2013, The Lancet. Oncology.

[39]  Takahiro Ochiya,et al.  The Roles of MicroRNAs in Breast Cancer , 2015, Cancers.

[40]  Nikitina Eg,et al.  MicroRNAs and human cancer. , 2012 .

[41]  A. Paradiso,et al.  Aberrant miRNAs expressed in HER-2 negative breast cancers patient , 2018, Journal of Experimental & Clinical Cancer Research.

[42]  Fei-Fei Liu,et al.  MicroRNA-320 suppresses colorectal cancer by targeting SOX4, FOXM1, and FOXQ1 , 2016, Oncotarget.

[43]  Fei-Fei Liu,et al.  Bone morphogenetic protein 2 (BMP2) induces growth suppression and enhances chemosensitivity of human colon cancer cells , 2016, Cancer Cell International.

[44]  T. Mcclanahan,et al.  Involvement of chemokine receptors in breast cancer metastasis , 2001, Nature.

[45]  C. Croce,et al.  MicroRNA-cancer connection: the beginning of a new tale. , 2006, Cancer research.

[46]  G. Goodall,et al.  The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1 , 2008, Nature Cell Biology.

[47]  Decoding the usefulness of non-coding RNAs as breast cancer markers , 2016, Journal of Translational Medicine.

[48]  Geoffrey Childs,et al.  Low-level expression of microRNAs let-7d and miR-205 are prognostic markers of head and neck squamous cell carcinoma. , 2009, The American journal of pathology.