KAI1 Gene Is Engaged in NDRG1 Gene-mediated Metastasis Suppression through the ATF3-NFκB Complex in Human Prostate Cancer*

NDRG1 and KAI1 belong to metastasis suppressor genes, which impede the dissemination of tumor cells from primary tumors to distant organs. Previously, we identified the metastasis promoting transcription factor, ATF3, as a downstream target of NDRG1. Further analysis revealed that the KAI1 promoter contained a consensus binding motif of ATF3, suggesting a possibility that NDRG1 suppresses metastasis through inhibition of ATF3 expression followed by activation of the KAI1 gene. In this report, we found that ectopic expression of NDRG1 was able to augment endogenous KAI1 gene expression in prostate cancer cell lines, whereas silencing NDRG1 was accompanied with significant decrease in KAI1 expression in vitro and in vivo. In addition, our results of ChIP analysis indicate that ATF3 indeed bound to the promoter of the KAI1 gene. Importantly, our promoter-based analysis revealed that ATF3 modulated KAI1 transcription through cooperation with other endogenous transcription factor as co-activator (ATF3-JunB) or co-repressor (ATF3-NFκB). Moreover, loss of KAI1 expression significantly abrogated NDRG1-mediated metastatic suppression in vitro as well as in a spontaneous metastasis animal model, indicating that KA11 is a functional downstream target of the NDRG1 pathway. Our result of immunohistochemical analysis showed that loss of NDRG1 and KAI1 occurs in parallel as prostate cancer progresses. We also found that a combined expression status of these two genes serves as a strong independent prognostic marker to predict metastasis-free survival of prostate cancer patients. Taken together, our result revealed a novel regulatory network of two metastasis suppressor genes, NDRG1 and KAI1, which together concerted metastasis-suppressive activities through an intrinsic transcriptional cascade.

[1]  Mitsutoshi Nakamura,et al.  Function of JunB in Transient Amplifying Cell Senescence and Progression of Human Prostate Cancer , 2008, Clinical Cancer Research.

[2]  Kathleen A. Kennedy,et al.  Systems biology approaches identify ATF3 as a negative regulator of Toll-like receptor 4 , 2006, Nature.

[3]  Tsonwin Hai,et al.  ATF3 and stress responses. , 1999, Gene expression.

[4]  K. Miura,et al.  The Drg-1 gene suppresses tumor metastasis in prostate cancer. , 2003, Cancer research.

[5]  T. Tsuruo,et al.  Identification of genes differentially expressed in B16 murine melanoma sublines with different metastatic potentials. , 1996, Cancer research.

[6]  Christopher K. Glass,et al.  Exchange of N-CoR Corepressor and Tip60 Coactivator Complexes Links Gene Expression by NF-κB and β-Amyloid Precursor Protein , 2002, Cell.

[7]  W. Dinjens,et al.  A novel gene which is up-regulated during colon epithelial cell differentiation and down-regulated in colorectal neoplasms. , 1997, Laboratory investigation; a journal of technical methods and pathology.

[8]  W. Isaacs,et al.  Genomic organization of the human KAI1 metastasis-suppressor gene. , 1997, Genomics.

[9]  P. Jackson,et al.  KAI1 promoter activity is dependent on p53, junB and AP2: evidence for a possible mechanism underlying loss of KAI1 expression in cancer cells , 2005, Oncogene.

[10]  P. Steeg,et al.  Metastasis suppressor genes: basic biology and potential clinical use. , 2003, Clinical breast cancer.

[11]  J. Barrett,et al.  Location of KAI1 on the short arm of human chromosome 11 and frequency of allelic loss in advanced human prostate cancer , 1997, The Prostate.

[12]  S. Clark,et al.  Methylation of a CpG island within the promoter region of the KAI1 metastasis suppressor gene is not responsible for down-regulation of KAI1 expression in invasive cancers or cancer cell lines. , 2000, Cancer letters.

[13]  K. Miura,et al.  The expression of the KAI1 gene, a tumor metastasis suppressor, is directly activated by p53. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[14]  R. Matusik,et al.  The nuclear factor-kappaB pathway controls the progression of prostate cancer to androgen-independent growth. , 2008, Cancer research.

[15]  C. Sheehan,et al.  Expression of nuclear factor-kappa B and I kappa B alpha proteins in prostatic adenocarcinomas: correlation of nuclear factor-kappa B immunoreactivity with disease recurrence. , 2004, Clinical cancer research : an official journal of the American Association for Cancer Research.

[16]  D. Welch,et al.  Metastasis suppressor pathways--an evolving paradigm. , 2003, Cancer letters.

[17]  V. Robinson,et al.  Metastasis suppressor genes: From gene identification to protein function and regulation , 2005, Cancer biology & therapy.

[18]  A. Rabson,et al.  NF‐κB activation in human prostate cancer: Important mediator or epiphenomenon? , 2004 .

[19]  The KISS1 metastasis suppressor: mechanistic insights and clinical utility. , 2006, Frontiers in bioscience : a journal and virtual library.

[20]  K. Kim,et al.  Expression of a splice variant of KAI1, a tumor metastasis suppressor gene, influences tumor invasion and progression. , 2003, Cancer research.

[21]  D. Richardson,et al.  The metastasis suppressor, Ndrg-1: a new ally in the fight against cancer. , 2006, Carcinogenesis.

[22]  Xiaoke Huang,et al.  Endoglin suppresses human prostate cancer metastasis , 2010, Clinical & Experimental Metastasis.

[23]  A. Scaloni,et al.  Transcription regulation by the adaptor protein Fe65 and the nucleosome assembly factor SET , 2005, EMBO reports.

[24]  T. Tsuruo,et al.  Inhibitory Effect of ATF3 Antisense Oligonucleotide on Ectopic Growth of HT29 Human Colon Cancer Cells , 2000, Japanese journal of cancer research : Gann.

[25]  W. Isaacs,et al.  Down-regulation of the KAI1 metastasis suppressor gene during the progression of human prostatic cancer infrequently involves gene mutation or allelic loss. , 1996, Cancer research.

[26]  H. Ford,et al.  Drg-1 as a differentiation-related, putative metastatic suppressor gene in human colon cancer. , 2000, Cancer research.

[27]  S. Eccles,et al.  Metastasis: recent discoveries and novel treatment strategies , 2007, The Lancet.

[28]  M. Kuwano,et al.  Tumor growth suppression in pancreatic cancer by a putative metastasis suppressor gene Cap43/NDRG1/Drg-1 through modulation of angiogenesis. , 2006, Cancer research.

[29]  P. Steeg.  Tumor metastasis: mechanistic insights and clinical challenges , 2006, Nature Medicine.

[30]  T. Nakajima,et al.  Mutation and expression of the metastasis suppressor gene KAI1 in esophageal squamous cell carcinoma , 2000, Cancer.

[31]  T. Kasamatsu,et al.  Cap43/NDRG1/Drg-1 is a molecular target for angiogenesis and a prognostic indicator in cervical adenocarcinoma. , 2008, Cancer letters.

[32]  P. Banerjee,et al.  Genistein induces the metastasis suppressor kangai-1 which mediates its anti-invasive effects in TRAMP cancer cells. , 2007, Biochemical and biophysical research communications.

[33]  K. Handa,et al.  Motility inhibition and apoptosis are induced by metastasis-suppressing gene product CD82 and its analogue CD9, with concurrent glycosylation. , 1999, Cancer research.

[34]  H. Izumi,et al.  N-myc downstream regulated gene 1/Cap43 suppresses tumor growth and angiogenesis of pancreatic cancer through attenuation of inhibitor of kappaB kinase beta expression. , 2009, Cancer research.

[35]  K. Watabe,et al.  Drug development against metastasis-related genes and their pathways: a rationale for cancer therapy. , 2008, Biochimica et biophysica acta.

[36]  D. Richardson,et al.  Iron Chelation by Clinically Relevant Anthracyclines: Alteration in Expression of Iron-Regulated Genes and Atypical Changes in Intracellular Iron Distribution and Trafficking , 2008, Molecular Pharmacology.

[37]  David A. Cheresh,et al.  Nuclear cytokine-activated IKKα controls prostate cancer metastasis by repressing Maspin , 2007, Nature.

[38]  J. Barrett,et al.  KAI1, a metastasis suppressor gene for prostate cancer on human chromosome 11p11.2. , 1995, Science.

[39]  Y. Takeshima,et al.  Down‐regulation of KAI1 Messenger RNA Expression Is Not Associated with Loss of Heterozygosity of the KAI1 Gene Region in Lung Adenocarcinoma , 1999, Japanese journal of cancer research : Gann.

[40]  Hee June Choi,et al.  Transcriptional regulation of a metastasis suppressor gene by Tip60 and β-catenin complexes , 2005, Nature.

[41]  M. Oshimura,et al.  Localization of metastasis suppressor gene(s) for prostatic cancer to the short arm of human chromosome 11. , 1992, Cancer research.

[42]  P. Jackson,et al.  Identification of regulatory regions within the KAI1 promoter: a role for binding of AP1, AP2 and p53. , 2003, Gene.

[43]  Ximing J. Yang,et al.  Nuclear Factor-κB-Mediated Transforming Growth Factor-β-Induced Expression of Vimentin Is an Independent Predictor of Biochemical Recurrence after Radical Prostatectomy , 2009, Clinical Cancer Research.

[44]  S. Grimm,et al.  The metastasis suppressor gene C33/CD82/KAI1 induces apoptosis through reactive oxygen intermediates , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[45]  Tsonwin Hai,et al.  The tumor metastasis suppressor gene Drg-1 down-regulates the expression of activating transcription factor 3 in prostate cancer. , 2006, Cancer research.

[46]  Wen-Lin Kuo,et al.  A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. , 2006, Cancer cell.

[47]  J. Isaacs,et al.  Genetic factors and suppression of metastatic ability of prostatic cancer. , 1991, Cancer research.

[48]  P. Steeg.  Metastasis suppressors alter the signal transduction of cancer cells , 2003, Nature Reviews Cancer.

[49]  T. Mashimo,et al.  Activation of the tumor metastasis suppressor gene, KAI1, by etoposide is mediated by p53 and c-Jun genes. , 2000, Biochemical and Biophysical Research Communications - BBRC.

[50]  Ping Zhang,et al.  NDRG1, a growth and cancer related gene: regulation of gene expression and function in normal and disease states. , 2007, Carcinogenesis.

[51]  Toshiki Watanabe,et al.  Differential usage of alternate promoters of the human stress response gene ATF3 in stress response and cancer cells , 2009, Nucleic acids research.

[52]  H. Gaugitsch,et al.  A new superfamily of lymphoid and melanoma cell proteins with extensive homology to Schistosoma mansoni antigen Sm23 , 1991, European journal of immunology.

[53]  S. Aaronson,et al.  Inhibition of tumor cell growth by RTP/rit42 and its responsiveness to p53 and DNA damage. , 1998, Cancer research.

[54]  Xin A. Zhang,et al.  KAI1/CD82, a tumor metastasis suppressor. , 2006, Cancer letters.