Downregulation of microRNA-182 inhibits cell growth and invasion by targeting programmed cell death 4 in human lung adenocarcinoma cells

Lung cancer is a major cause of cancer death worldwide. Programmed cell death 4 (PDCD4), an important tumor suppressor, influences transcription and translation of multiple genes and modulates different signal transduction pathways. However, the upstream regulation of this gene is largely unknown. In our study, we found that microRNA-182 (miR-182) was upregulated, whereas PDCD4 was downregulated in lung cancer cell lines. We performed methyl thiazolyl tetrazolium and colony formation assays to study the influence of miR-182 on proliferation of the lung cancer cell lines A549 and SPC-A-1. We also carried out Transwell and wound healing assays to investigate the effect of miR-182 on invasion and migration of A549 and SPC-A-1. Finally, using the luciferase reporter assay and restore assay, we demonstrated that PDCD4 is a direct target of miR-182. These results suggest that in lung adenocarcinoma cells, miR-182 plays an oncogenic role as a direct negative regulator of PDCD4.

[1]  Phillip D. Zamore,et al.  Ribo-gnome: The Big World of Small RNAs , 2005, Science.

[2]  Jin-Wu Nam,et al.  Genomics of microRNA. , 2006, Trends in genetics : TIG.

[3]  Zijie Sun,et al.  Downregulation of tumor suppressor Pdcd4 promotes invasion and activates both β-catenin/Tcf and AP-1-dependent transcription in colon carcinoma cells , 2008, Oncogene.

[4]  N. Colburn,et al.  Tumorigenesis Suppressor Pdcd4 Down-Regulates Mitogen-Activated Protein Kinase Kinase Kinase Kinase 1 Expression To Suppress Colon Carcinoma Cell Invasion , 2006, Molecular and Cellular Biology.

[5]  Rende Guo,et al.  MicroRNA‐182 promotes cell growth, invasion, and chemoresistance by targeting programmed cell death 4 (PDCD4) in human ovarian carcinomas , 2013, Journal of cellular biochemistry.

[6]  C. Burge,et al.  Conserved Seed Pairing, Often Flanked by Adenosines, Indicates that Thousands of Human Genes are MicroRNA Targets , 2005, Cell.

[7]  C. Croce,et al.  MicroRNA gene expression deregulation in human breast cancer. , 2005, Cancer research.

[8]  C. Fujiyama,et al.  Novel human PDCD4 (H731) gene expressed in proliferative cells is expressed in the small duct epithelial cells of the breast as revealed by an anti‐H731 antibody , 1999, Pathology international.

[9]  Nahum Sonenberg,et al.  A Novel Function of the MA-3 Domains in Transformation and Translation Suppressor Pdcd4 Is Essential for Its Binding to Eukaryotic Translation Initiation Factor 4A , 2004, Molecular and Cellular Biology.

[10]  R. Stephens,et al.  Unique microRNA molecular profiles in lung cancer diagnosis and prognosis. , 2006, Cancer cell.

[11]  Hiroshi I. Suzuki,et al.  Modulation of microRNA processing by p53 , 2009, Nature.

[12]  C. Croce,et al.  MicroRNA signatures in human cancers , 2006, Nature Reviews Cancer.

[13]  A. Jemal,et al.  Cancer Statistics, 2010 , 2010, CA: a cancer journal for clinicians.

[14]  T. Honjo,et al.  Isolation of a novel mouse gene MA-3 that is induced upon programmed cell death. , 1995, Gene.

[15]  V. Ambros,et al.  The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14 , 1993, Cell.

[16]  D. Ettinger,et al.  Multidisciplinary management of lung cancer. , 2004, The New England journal of medicine.

[17]  Min Liu,et al.  MicroRNA‐182 targets cAMP‐responsive element‐binding protein 1 and suppresses cell growth in human gastric adenocarcinoma , 2012, The FEBS journal.

[18]  A. Krogh,et al.  Programmed Cell Death 4 (PDCD4) Is an Important Functional Target of the MicroRNA miR-21 in Breast Cancer Cells* , 2008, Journal of Biological Chemistry.

[19]  Na Li,et al.  The microRNA miR-182 is induced by IL-2 and promotes clonal expansion of activated helper T lymphocytes , 2010, Nature Immunology.

[20]  C. Croce,et al.  The role of microRNA genes in papillary thyroid carcinoma. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Julius Brennecke,et al.  Towards a complete description of the microRNA complement of animal genomes , 2003, Genome Biology.

[22]  N. Colburn,et al.  Programmed Cell Death 4 inhibits breast cancer cell invasion by increasing Tissue Inhibitor of Metalloproteinases-2 expression , 2009, Breast Cancer Research and Treatment.

[23]  Fang Zhou,et al.  Distinctive microRNA profiles relating to patient survival in esophageal squamous cell carcinoma. , 2008, Cancer research.

[24]  L. Tanoue Cancer Statistics, 2009 , 2010 .

[25]  D. Bartel MicroRNAs Genomics, Biogenesis, Mechanism, and Function , 2004, Cell.

[26]  Y. Tomari,et al.  The microRNA pathway and cancer , 2010, Cancer science.

[27]  R. Göke,et al.  The tumour suppressor Pdcd4: recent advances in the elucidation of function and regulation , 2009, Biology of the cell.

[28]  D. Polsky,et al.  Aberrant miR-182 expression promotes melanoma metastasis by repressing FOXO3 and microphthalmia-associated transcription factor , 2009, Proceedings of the National Academy of Sciences.

[29]  V. Ambros The functions of animal microRNAs , 2004, Nature.

[30]  Yun Zhang,et al.  PDCD4 gene silencing in gliomas is associated with 5′CpG island methylation and unfavourable prognosis , 2008, Journal of cellular and molecular medicine.

[31]  George A Calin,et al.  MicroRNA expression profiles associated with prognosis and therapeutic outcome in colon adenocarcinoma. , 2008, JAMA.

[32]  C. Croce,et al.  MicroRNA signatures in human ovarian cancer. , 2007, Cancer research.

[33]  H. Allgayer,et al.  MicroRNA-21 (miR-21) post-transcriptionally downregulates tumor suppressor Pdcd4 and stimulates invasion, intravasation and metastasis in colorectal cancer , 2008, Oncogene.

[34]  N. Colburn,et al.  Differentially expressed protein Pdcd4 inhibits tumor promoter-induced neoplastic transformation. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[35]  K. Uematsu,et al.  Genomic alterations in human mesothelioma including high resolution mapping of common regions of DNA loss in chromosome arm 6q. , 2003, Anticancer research.

[36]  N. Dubrawsky Cancer statistics , 1989, CA: a cancer journal for clinicians.

[37]  N. Colburn,et al.  Epidermal expression of the translation inhibitor programmed cell death 4 suppresses tumorigenesis. , 2005, Cancer research.

[38]  G. Ruvkun,et al.  Posttranscriptional regulation of the heterochronic gene lin-14 by lin-4 mediates temporal pattern formation in C. elegans , 1993, Cell.

[39]  Zhaojian Liu,et al.  MiR‐182 overexpression in tumourigenesis of high‐grade serous ovarian carcinoma , 2012, The Journal of pathology.

[40]  A. Yoshida,et al.  MicroRNA-21 correlates with tumorigenesis in malignant peripheral nerve sheath tumor (MPNST) via programmed cell death protein 4 (PDCD4) , 2012, Journal of Cancer Research and Clinical Oncology.

[41]  A. Jemal,et al.  Cancer Statistics, 2009 , 2009, CA: a cancer journal for clinicians.

[42]  Sun Mi Park,et al.  MicroRNAs: key players in the immune system, differentiation, tumorigenesis and cell death , 2008, Oncogene.

[43]  K. Kosik,et al.  MicroRNA-21 is an antiapoptotic factor in human glioblastoma cells. , 2005, Cancer research.

[44]  A. Wlodawer,et al.  Mutational analysis of the DEAD-box RNA helicase eIF4AII characterizes its interaction with transformation suppressor Pdcd4 and eIF4GI. , 2005, RNA.

[45]  Thomas Tuschl,et al.  miRNAs in human cancer , 2011, The Journal of pathology.