Drastic down-regulation of Krüppel-like factor 4 expression is critical in human gastric cancer development and progression.

Krüppel-like factor 4 (KLF4) is highly expressed in epithelial tissues such as the gut and skin. However, the role of KLF4 in human gastric cancer development and progression is unknown. Here we show that KLF4 protein expression was decreased or lost in primary tumors and, in particular, lymph node metastases when compared with that in normal gastric mucosa. Moreover, loss of KLF4 expression in the primary tumors was significantly associated with poor survival, and also an independent prognostic marker in a multivariate analysis. Consistently, most human gastric cancer cell lines exhibited loss of or a substantial decrease in KLF4 expression at both RNA and protein levels. Enforced restoration of KLF4 expression resulted in marked cell growth inhibition in vitro and significantly attenuated tumor growth and total abrogation of metastasis in an orthotopic animal model of gastric cancer. Mechanism studies indicated that promoter hypermethylation and hemizygous deletion contributed to the down-regulation of KLF4 expression and the induction of apoptosis contributed to the antitumor activity of KLF4. Collectively, our data provide first clinical and casual evidence and potential mechanism that the alteration of KLF4 expression plays a critical role in gastric cancer development and progression.

[1]  N. Wright,et al.  The gastrointestinal stem cell , 2004, Cell proliferation.

[2]  Channing S. Mahatan,et al.  The Gut-enriched Krüppel-like Factor (Krüppel-like Factor 4) Mediates the Transactivating Effect of p53 on the p21 WAF1/Cip1 Promoter* , 2000, The Journal of Biological Chemistry.

[3]  N. Wright,et al.  Gastrointestinal stem cells , 2002, The Journal of pathology.

[4]  N. Wright,et al.  INTRAVENOUS EPIDERMAL GROWTH FACTOR/UROGASTRONE INCREASES SMALL-INTESTINAL CELL PROLIFERATION IN CONGENITAL MICROVILLOUS ATROPHY , 1985, The Lancet.

[5]  S. I. Bae,et al.  Genetic alterations in gastric cancer cell lines and their original tissues , 2000, International journal of cancer.

[6]  Zhihua Liu,et al.  Discovery of Ca2+-relevant and differentiation-associated genes downregulated in esophageal squamous cell carcinoma using cDNA microarray , 2004, Oncogene.

[7]  K. Kaestner,et al.  Foxl1 Controls the Wnt/β-Catenin Pathway by Modulating the Expression of Proteoglycans in the Gut* , 2001, The Journal of Biological Chemistry.

[8]  C. Moskaluk,et al.  Inactivation of the E-Cadherin Gene in Sporadic Diffuse-Type Gastric Cancer , 2001, Modern Pathology.

[9]  Hall,et al.  Gastrointestinal Pathology , 1990, Current Topics in Pathology.

[10]  J. Abbruzzese,et al.  Celecoxib Inhibits Vascular Endothelial Growth Factor Expression in and Reduces Angiogenesis and Metastasis of Human Pancreatic Cancer via Suppression of Sp1 Transcription Factor Activity , 2004, Cancer Research.

[11]  A. Majumdar,et al.  Regulation of gastrointestinal mucosal growth during aging. , 2003, Journal of physiology and pharmacology : an official journal of the Polish Physiological Society.

[12]  Jun Yu,et al.  Increased β-catenin mRNA levels and mutational alterations of the APC and β-catenin gene are present in intestinal-type gastric cancer , 2002 .

[13]  Ph.D. Eiichi Tahara M.D.,et al.  Molecular biology of gastric cancer , 2004, World Journal of Surgery.

[14]  V. Yang,et al.  Requirement of Krüppel-like Factor 4 in Preventing Entry into Mitosis following DNA Damage* , 2004, Journal of Biological Chemistry.

[15]  H. Höfler,et al.  Relationship between E‐cadherin gene mutation and p53 gene mutation, p53 accumulation, Bcl‐2 expression and Ki‐67 staining in diffuse‐type gastric carcinoma , 2003, International journal of cancer.

[16]  D. Fang,et al.  Mutation analysis of APC gene in gastric cancer with microsatellite instability. , 2002, World journal of gastroenterology.

[17]  Study of the repairing process of gastric ulcer using multivariate analysis of bFGF-positive cells, haemodynamics, PAS-positive mucus amount and glandular index in the gastric mucosa. , 1996, Journal of gastroenterology and hepatology.

[18]  T. Akiyama,et al.  Candidate regions of tumor suppressor locus on chromosome 9q31.1 in gastric cancer , 2004, International journal of cancer.

[19]  J. Ruppert,et al.  Increase of GKLF messenger RNA and protein expression during progression of breast cancer. , 2000, Cancer research.

[20]  J. Luk,et al.  Molecular biology of gastric carcinoma: From laboratory to bedside , 1999, Journal of gastroenterology and hepatology.

[21]  A. Jemal,et al.  Global cancer statistics , 2011, CA: a cancer journal for clinicians.

[22]  C. Stone,et al.  Gut‐enriched Krüppel‐like factor regulates colonic cell growth through APC/β‐catenin pathway , 2002, FEBS letters.

[23]  V. Yang,et al.  Transcriptional profiling of Krüppel-like factor 4 reveals a function in cell cycle regulation and epithelial differentiation. , 2003, Journal of molecular biology.

[24]  M. Tomikawa,et al.  Gastrointestinal mucosal regeneration: role of growth factors. , 1999, Frontiers in bioscience : a journal and virtual library.

[25]  I. Talbot,et al.  Genetic alterations in gastric cancers from British patients. , 2001, Cancer genetics and cytogenetics.

[26]  M. Matsuoka,et al.  Identification of Aberrantly Methylated Genes in Association with Adult T-Cell Leukemia , 2004, Cancer Research.

[27]  J. M. Shields,et al.  Identification and Characterization of a Gene Encoding a Gut-enriched Krüppel-like Factor Expressed during Growth Arrest* , 1996, The Journal of Biological Chemistry.

[28]  J. Ajani,et al.  Transcription factor Sp1 expression is a significant predictor of survival in human gastric cancer. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[29]  E. Marbán,et al.  Krüppel-like Factor 4 (Gut-enriched Krüppel-like Factor) Inhibits Cell Proliferation by Blocking G1/S Progression of the Cell Cycle* , 2001, The Journal of Biological Chemistry.

[30]  Channing S. Mahatan,et al.  Expression of the gut-enriched Krüppel-like factor (Krüppel-like factor 4) gene in the human colon cancer cell line RKO is dependent on CDX2 , 2001, Oncogene.

[31]  V. Yang,et al.  Identification of Krüppel-like factor 4 as a potential tumor suppressor gene in colorectal cancer , 2004, Oncogene.

[32]  A. Tarnawski,et al.  Regeneration of gastric mucosa during ulcer healing is triggered by growth factors and signal transduction pathways , 2001, Journal of Physiology-Paris.

[33]  K. Kaestner,et al.  The mesenchymal winged helix transcription factor Fkh6 is required for the control of gastrointestinal proliferation and differentiation. , 1997, Genes & development.

[34]  R. P. Blankfield,et al.  Helicobacter pylori infection and the development of gastric cancer. , 2001, The New England journal of medicine.

[35]  Min Wu,et al.  Down-regulation of gut-enriched Kruppel-like factor expression in esophageal cancer. , 2002, World journal of gastroenterology.

[36]  Michael F. Seldin,et al.  A Gene for a Novel Zinc-finger Protein Expressed in Differentiated Epithelial Cells and Transiently in Certain Mesenchymal Cells* , 1996, The Journal of Biological Chemistry.

[37]  Teruhiko Yoshida,et al.  Downregulation and growth inhibitory effect of epithelial-type Krüppel-like transcription factor KLF4, but not KLF5, in bladder cancer. , 2003, Biochemical and biophysical research communications.

[38]  D. Roberts,et al.  Sonic hedgehog expression correlates with fundic gland differentiation in the adult gastrointestinal tract , 2002, Gut.

[39]  E. Kay,et al.  APC Gene Expression in Gastric Carcinoma: An Immunohistochemical Study , 2002, Applied immunohistochemistry & molecular morphology : AIMM.

[40]  E. Tahara Molecular aspects of invasion and metastasis of stomach cancer. , 2000, Verhandlungen der Deutschen Gesellschaft fur Pathologie.

[41]  J. Gordon,et al.  Differentiation and self-renewal in the mouse gastrointestinal epithelium. , 1994, Current opinion in cell biology.

[42]  Jacques Ferlay,et al.  Estimates of the worldwide incidence of 25 major cancers in 1990 , 1999, International journal of cancer.

[43]  J. Shie,et al.  Up‐regulation of gut‐enriched krüppel‐like factor by interferon‐γ in human colon carcinoma cells , 2000 .

[44]  V. Yang,et al.  Krüppel-like Factor 4 Mediates p53-dependent G1/S Cell Cycle Arrest in Response to DNA Damage* , 2003, The Journal of Biological Chemistry.

[45]  J. Ruppert,et al.  Oncogene expression cloning by retroviral transduction of adenovirus E1A-immortalized rat kidney RK3E cells: transformation of a host with epithelial features by c-MYC and the zinc finger protein GKLF. , 1999, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[46]  C. Fenoglio-Preiser,et al.  TP53 and gastric carcinoma: A review , 2003, Human mutation.

[47]  Z. Y. Chen,et al.  Gut-enriched Krüppel-like factor represses cyclin D1 promoter activity through Sp1 motif. , 2000, Nucleic acids research.