Oncogenic function of the homeobox A13-long noncoding RNA HOTTIP-insulin growth factor-binding protein 3 axis in human gastric cancer

To study the mechanisms of gastric tumorigenesis, we have established CSN cell line from human normal gastric mucosa, and CS12, a tumorigenic and invasive gastric cancer cell line from CSN passages. Many stem cell markers were expressed in both CSN and CS12 cells, but LGR5 and NANOG were expressed only in CS12 cells. Increased expression of homeobox A13 (HoxA13) and its downstream cascades was significant for the tumorigenic activity of CS12 cells, and was associated with recruitment of E2F-1 to HoxA13 promoter accompanied with increased trimethylation of histone H3 lysine 4 (H3K4me3) at the hypomethylated E2F motifs. Knockdown of HoxA13 caused the downregulation of long non-coding RNA HOTTIP and insulin growth factor-binding protein 3 (IGFBP-3) genes, indicating that both were targets of HoxA13. Concurrent regulation of HoxA13-HOTTIP was mediated by the mixed lineage leukemia-WD repeat domain 5 complex, which caused the trimethylation of H3K4 and then stimulated cell proliferation. HoxA13 transactivated the IGFBP-3 promoter through the HOX-binding site. Activation of IGFBP-3 stimulated the oncogenic potential and invasion activity. Increased expression of HoxA13 (63.2%) and IGFBP-3 (28.6%) was detected in human gastric cancer tissues and was found in the gastric cancer data of The Cancer Genome Atlas. Taken together, the HoxA13–HOTTIP–IGFBP-3 cascade is critical for the carcinogenic characteristics of CS12 cells.

[1]  A. Jemal,et al.  Cancer Statistics, 2006 , 2006, CA: a cancer journal for clinicians.

[2]  Sebastian Böcker,et al.  High-throughput MALDI-TOF discovery of genomic sequence polymorphisms. , 2003, Genome research.

[3]  C. Tickle,et al.  HOX-4 genes and the morphogenesis of mammalian genitalia. , 1991, Genes & development.

[4]  D. Mortlock,et al.  The molecular basis of hypodactyly (Hd): a deletion in Hoxa13 leads to arrest of digital arch formation , 1996, Nature Genetics.

[5]  D. Mortlock,et al.  Mutation of HOXA13 in hand-foot-genital syndrome , 1997, Nature Genetics.

[6]  Dean Y. Li,et al.  HOXA13 Is Essential for Placental Vascular Patterning and Labyrinth Endothelial Specification , 2008, PLoS genetics.

[7]  Yinxue Wang,et al.  The long non-coding RNA HOTTIP promotes progression and gemcitabine resistance by regulating HOXA13 in pancreatic cancer , 2015, Journal of Translational Medicine.

[8]  Y. Ko,et al.  Androgen receptor-mediated apoptosis in bovine testicular induced pluripotent stem cells in response to phthalate esters , 2013, Cell Death and Disease.

[9]  D. Hanahan,et al.  The Hallmarks of Cancer , 2000, Cell.

[10]  J. Corton,et al.  The long non-coding RNA HOTTIP enhances pancreatic cancer cell proliferation, survival and migration , 2015, Oncotarget.

[11]  F. Roviello,et al.  Multimodal treatment of gastric cancer in the west: Where are we going? , 2015, World journal of gastroenterology.

[12]  S. Firth,et al.  IGFBP-3: a cell fate pivot in cancer and disease. , 2014, Growth hormone & IGF research : official journal of the Growth Hormone Research Society and the International IGF Research Society.

[13]  Andrew G. Clark,et al.  Modes of cancer cell invasion and the role of the microenvironment. , 2015, Current opinion in cell biology.

[14]  S. Mundlos,et al.  Polyalanine expansion in HOXA13: three new affected families and the molecular consequences in a mouse model. , 2004, Human molecular genetics.

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

[16]  H. Schöler,et al.  OCT4: dynamic DNA binding pioneers stem cell pluripotency. , 2014, Biochimica et biophysica acta.

[17]  Ling Huang,et al.  5-Fluorouracil Chemotherapy of Gastric Cancer Generates Residual Cells with Properties of Cancer Stem Cells , 2015, International journal of biological sciences.

[18]  S. Velasco-Miguel,et al.  Induction of the growth inhibitor IGF-binding protein 3 by p53 , 1995, Nature.

[19]  C. Sherr.  Cancer Cell Cycles , 1996, Science.

[20]  S. Shou,et al.  Survival of Hoxa13 homozygous mutants reveals a novel role in digit patterning and appendicular skeletal development , 2010, Developmental dynamics : an official publication of the American Association of Anatomists.

[21]  C. Jin,et al.  Jun dimerization protein 2 is a critical component of the Nrf2/MafK complex regulating the response to ROS homeostasis , 2013, Cell Death and Disease.

[22]  J. Schwander,et al.  Insulin-Like Growth Factor-Binding Protein , 1989, Definitions.

[23]  M. Davey,et al.  Urea improves efficiency of bisulphite-mediated sequencing of 5'-methylcytosine in genomic DNA. , 1998, Nucleic acids research.

[24]  Saraswati Sukumar,et al.  The Hox genes and their roles in oncogenesis , 2010, Nature Reviews Cancer.

[25]  R. Baxter.  Nuclear actions of insulin-like growth factor binding protein-3. , 2015, Gene.

[26]  P Chambon,et al.  Hoxa-13 and Hoxd-13 play a crucial role in the patterning of the limb autopod. , 1996, Development.

[27]  Ke-Neng Chen,et al.  Exploration of target genes of HOXA13 in esophageal squamous cell carcinoma cell line. , 2011, Cancer letters.

[28]  John K Field,et al.  Quantitative high-throughput analysis of DNA methylation patterns by base-specific cleavage and mass spectrometry. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[29]  Howard Y. Chang,et al.  A long noncoding RNA maintains active chromatin to coordinate homeotic gene expression , 2011, Nature.

[30]  A. Kuroiwa,et al.  Coordinated expression of Abd-B subfamily genes of the HoxA cluster in the developing digestive tract of chick embryo. , 1995, Developmental biology.

[31]  Chia‐cheng Chang,et al.  Isolation and characterization of human gastric cell lines with stem cell phenotypes , 2007, Journal of gastroenterology and hepatology.

[32]  Yong Li,et al.  HOXA13 promotes cancer cell growth and predicts poor survival of patients with esophageal squamous cell carcinoma. , 2009, Cancer research.

[33]  A. Yoshiki,et al.  Suppression of cell-cycle progression by Jun dimerization protein-2 (JDP2) involves downregulation of cyclin-A2 , 2009, Oncogene.

[34]  Howard Y. Chang,et al.  Functional Demarcation of Active and Silent Chromatin Domains in Human HOX Loci by Noncoding RNAs , 2007, Cell.

[35]  K M Higgins,et al.  Loss of Eph-receptor expression correlates with loss of cell adhesion and chondrogenic capacity in Hoxa13 mutant limbs. , 2001, Development.

[36]  H. Shiraha,et al.  Functional promoter upstream p53 regulatory sequence of IGFBP3 that is silenced by tumor specific methylation , 2005, BMC Cancer.

[37]  A. Guttmacher.  Autosomal dominant preaxial deficiency, postaxial polydactyly, and hypospadias. , 1993, American journal of medical genetics.

[38]  N. Jones,et al.  p300 and ATF-2 are components of the DRF complex, which regulates retinoic acid- and E1A-mediated transcription of the c-jun gene in F9 cells. , 1998, Genes & development.

[39]  J. Zucman‐Rossi,et al.  The HOX gene network in hepatocellular carcinoma , 2011, International journal of cancer.

[40]  M. Xi,et al.  Long non-coding RNA HOTTIP is correlated with progression and prognosis in tongue squamous cell carcinoma , 2015, Tumor Biology.

[41]  K. Itakura,et al.  Expression of the Hoxa-13 gene correlates to hepatitis B and C virus associated HCC. , 2001, Biochemical and biophysical research communications.

[42]  M. Moses,et al.  Matrix metalloproteinases as novel biomarkers and potential therapeutic targets in human cancer. , 2009, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[43]  S. Kasper.  Exploring the Origins of the Normal Prostate and Prostate Cancer Stem Cell , 2008, Stem Cell Reviews.

[44]  Yang Han,et al.  Identification and validation that up-expression of HOXA13 is a novel independent prognostic marker of a worse outcome in gastric cancer based on immunohistochemistry , 2013, Medical Oncology.

[45]  P. Scambler,et al.  Novel HOXA13 mutations and the phenotypic spectrum of hand-foot-genital syndrome. , 2000, American journal of human genetics.

[46]  Howard Y. Chang,et al.  Long noncoding RNAs in cell-fate programming and reprogramming. , 2014, Cell stem cell.

[47]  Christian Ruiz,et al.  Long noncoding RNA HOTTIP/HOXA13 expression is associated with disease progression and predicts outcome in hepatocellular carcinoma patients , 2014, Hepatology.

[48]  D. Livingston,et al.  Distinct roles of the co-activators p300 and CBP in retinoic-acid-induced F9-cell differentiation , 1998, Nature.

[49]  J. Holly,et al.  Epithelial-to-mesenchymal transition in breast cancer: a role for insulin-like growth factor I and insulin-like growth factor–binding protein 3? , 2015, Breast cancer.