Aspirin Inhibits IKK-β-mediated Prostate Cancer Cell Invasion by Targeting Matrix Metalloproteinase-9 and Urokinase-Type Plasminogen Activator

Background/Aims: Aspirin has been demonstrated to possess potent chemopreventive and anticancer effects on prostate cancer. However, the more detailed molecular mechanisms of aspirin to suppress prostate cancer cell invasion have not been clearly elucidated. Methods: Transwell assays were performed to evaluate the effects of aspirin on cell invasion. Matrix metalloproteinases (MMPs) and serine proteinases activities in cell media were examined by gelatin zymography and ELISA. In addition, inhibitor of κB (IκB) kinase-β (IKK-β) phosphorylation and IKK-β kinase activity were measured to assess the effects of aspirin on IKK-β activation. Results: We found that aspirin suppressed the invasion and attachment in human prostate cancer cells. Aspirin treatment significantly resulted in reduction of matrix metalloproteinase-9 (MMP-9) and upregulation of tissue inhibitors of metalloproteinase-1 (TIMP-1) activity, which are the proteolytic enzymes contributing to the degradation of extracellular matrix and basement membrane in cell invasion and metastasis. Our data further showed that aspirin was able to inhibit both urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) expression in the cells. In addition, aspirin treatment caused a strong decrease in nuclear factor-kappa B (NF-κB) activation, inhibitor of κB (IκB)-α phosphorylation together with translocation of NF-κB p65 to nucleus and IκB kinase (IKK)- β activation. Moreover, the inhibitory effects of aspirin on cell invasion were reversed by IKK-β overexpression, while the IKK inhibitor sensitizes the anti-invasive effect of aspirin in prostate cancer cells. Conclusion: The present research concluded that aspirin suppressed prostate cancer cell invasion by reducing MMP-9 activity and uPA expression through decreasing of IKK-β-mediated NF-κB activation, indicating that the ability of aspirin to inhibit cell invasion might be useful in the chemoprevention of metastatic prostate cancer.

[1]  C. Brinckerhoff,et al.  The AP-1 site and MMP gene regulation: what is all the fuss about? , 1997, Matrix biology : journal of the International Society for Matrix Biology.

[2]  M. Skobe,et al.  Absence of host plasminogen activator inhibitor 1 prevents cancer invasion and vascularization , 1998, Nature Medicine.

[3]  D. Loskutoff,et al.  Regulation of cell adhesion by PAI‐1 , 1999, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[4]  J. Fata,et al.  Host TIMP-1 overexpression confers resistance to experimental brain metastasis of a fibrosarcoma cell line , 1998, Oncogene.

[5]  G. Andriole,et al.  Aspirin, NSAIDs, and Risk of Prostate Cancer: Results from the REDUCE Study , 2014, Clinical Cancer Research.

[6]  J. Christman,et al.  Expression by Direct Interaction with I B Activation and Cyclooxygenase-2 Κ Nf- Hepatitis C Virus Core Protein Suppresses , 2004 .

[7]  T. Uchiumi,et al.  Prostaglandin receptor EP3 mediates growth inhibitory effect of aspirin through androgen receptor and contributes to castration resistance in prostate cancer cells. , 2013, Endocrine-related cancer.

[8]  S. Devesa,et al.  International trends and patterns of prostate cancer incidence and mortality , 2000, International journal of cancer.

[9]  W. Zhong,et al.  CD147, MMP-1, MMP-2 and MMP-9 Protein Expression as Significant Prognostic Factors in Human Prostate Cancer , 2008, Oncology.

[10]  B. Binder,et al.  Urokinase-type plasminogen activator as a marker for the formation of distant metastases in prostatic carcinomas. , 1988, The Journal of urology.

[11]  A. Nalla,et al.  Targeting MMP-9, uPAR, and cathepsin B inhibits invasion, migration and activates apoptosis in prostate cancer cells , 2010, Cancer Gene Therapy.

[12]  M. Thun,et al.  Aspirin and non-steroidal anti-inflammatory drugs for cancer prevention: an international consensus statement. , 2009, The Lancet. Oncology.

[13]  B. Liu,et al.  Acetylbritannilactone Inhibits Neointimal Hyperplasia after Balloon Injury of Rat Artery by Suppressing Nuclear Factor-κB Activation , 2008, Journal of Pharmacology and Experimental Therapeutics.

[14]  T. Salo,et al.  Proteolytic degradation of extracellular matrix in tumor invasion. , 1987, Biochimica et biophysica acta.

[15]  G. Haas,et al.  The worldwide epidemiology of prostate cancer: perspectives from autopsy studies. , 2008, The Canadian journal of urology.

[16]  J. Christman,et al.  Hepatitis C virus core protein suppresses NF-kappaB activation and cyclooxygenase-2 expression by direct interaction with IkappaB kinase beta. , 2005, Journal of virology.

[17]  M. Luciani,et al.  Aspirin blocks proliferation in colon cells by inducing a G1 arrest and apoptosis through activation of the checkpoint kinase ATM. , 2007, Carcinogenesis.

[18]  S. Babykutty,et al.  Nimbolide retards tumor cell migration, invasion, and angiogenesis by downregulating MMP‐2/9 expression via inhibiting ERK1/2 and reducing DNA‐binding activity of NF‐κB in colon cancer cells , 2012, Molecular carcinogenesis.

[19]  R. Westrick,et al.  Plasminogen Activator Inhibitor-1 Regulates Tumor Growth and Angiogenesis* , 2001, The Journal of Biological Chemistry.

[20]  R. Everson,et al.  Aspirin Use and Lung, Colon, and Breast Cancer Incidence in a Prospective Study , 1994, Epidemiology.

[21]  R. Ball,et al.  Identification of degradome components associated with prostate cancer progression by expression analysis of human prostatic tissues , 2005, British Journal of Cancer.

[22]  K. Behbehani,et al.  TLR2 and AP-1/NF-kappaB are involved in the regulation of MMP-9 elicited by heat killed Listeria monocytogenes in human monocytic THP-1 cells , 2015, Journal of Inflammation.

[23]  W. Folk,et al.  Expression of urokinase plasminogen activator and receptor in conjunction with the ets family and AP-1 complex transcription factors in high grade prostate cancers. , 2001, European journal of cancer.

[24]  Zhiwei Wang,et al.  Inhibition of angiogenesis and invasion by 3,3'-diindolylmethane is mediated by the nuclear factor-kappaB downstream target genes MMP-9 and uPA that regulated bioavailability of vascular endothelial growth factor in prostate cancer. , 2007, Cancer research.

[25]  M. Rath,et al.  Patterns of MMP-2 and MMP-9 expression in human cancer cell lines. , 2009, Oncology reports.

[26]  Y. Shih,et al.  Antimetastatic potential of fisetin involves inactivation of the PI3K/Akt and JNK signaling pathways with downregulation of MMP-2/9 expressions in prostate cancer PC-3 cells , 2009, Molecular and Cellular Biochemistry.

[27]  S. Katiyar,et al.  Treatment of epigallocatechin‐3‐gallate inhibits matrix metalloproteinases‐2 and ‐9 via inhibition of activation of mitogen‐activated protein kinases, c‐jun and NF‐κB in human prostate carcinoma DU‐145 cells , 2004, The Prostate.

[28]  R. Bergan,et al.  Genistein inhibits human prostate cancer cell detachment, invasion, and metastasis. , 2014, The American journal of clinical nutrition.

[29]  K. Hu,et al.  Aspirin Suppresses the Growth and Metastasis of Osteosarcoma through the NF-κB Pathway , 2015, Clinical Cancer Research.

[30]  B. Liu,et al.  Acetylbritannilactone induces G1 arrest and apoptosis in vascular smooth muscle cells. , 2011, International journal of cardiology.

[31]  P. Andreasen,et al.  The plasminogen activation system in tumor growth, invasion, and metastasis , 2000, Cellular and Molecular Life Sciences CMLS.

[32]  D. Sliva,et al.  Aspirin inhibits highly invasive prostate cancer cells. , 2003, International journal of oncology.

[33]  C. Perez-stable,et al.  NF-κB activation enhances cell death by antimitotic drugs in human prostate cancer cells , 2010, Molecular Cancer.

[34]  S. Hawthorne,et al.  Amplification of MMP‐2 and MMP‐9 production by prostate cancer cell lines via activation of protease‐activated receptors , 2004, The Prostate.

[35]  M. Duffy,et al.  Proteases as prognostic markers in cancer. , 1996, Clinical cancer research : an official journal of the American Association for Cancer Research.

[36]  C. Evans,et al.  NF-kappaB 2 / p 52 induces resistance to Enzalutamide in Prostate Cancer : Role of androgen receptor and its variants , 2013 .

[37]  Ravikumar Aalinkeel,et al.  Gene Expression of Angiogenic Factors Correlates with Metastatic Potential of Prostate Cancer Cells , 2004, Cancer Research.

[38]  S. Lim,et al.  Isoliquiritigenin inhibits migration and invasion of prostate cancer cells: possible mediation by decreased JNK/AP-1 signaling. , 2009, The Journal of nutritional biochemistry.

[39]  Shigeyoshi Itohara,et al.  Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis , 2000, Nature Cell Biology.

[40]  B. Fingleton,et al.  Matrix metalloproteinases: biologic activity and clinical implications. , 2000, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[41]  S. Rabbani,et al.  Role of urokinase (uPA) and its receptor (uPAR) in invasion and metastasis of hormone-dependent malignancies. , 1998, International journal of oncology.

[42]  S. Karger,et al.  Cellular Physiology and Biochemistry Cellular Physiology and Biochemistry , 2021 .

[43]  S. Freedland,et al.  Daily aspirin use and prostate cancer-specific mortality in a large cohort of men with nonmetastatic prostate cancer. , 2014, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[44]  T. Deguchi,et al.  CCR1/CCL5 interaction promotes invasion of taxane-resistant PC3 prostate cancer cells by increasing secretion of MMPs 2/9 and by activating ERK and Rac signaling. , 2013, Cytokine.

[45]  K. Kerlikowske,et al.  Protective association of aspirin/NSAIDs and esophageal cancer: a systematic review and meta-analysis. , 2003, Gastroenterology.

[46]  M. Duffy,et al.  The urokinase‐type plasminogen activator system in cancer metastasis: A review , 1997, International journal of cancer.

[47]  C. Gondi,et al.  RNA interference-directed knockdown of urokinase plasminogen activator and urokinase plasminogen activator receptor inhibits prostate cancer cell invasion, survival, and tumorigenicity in vivo. , 2006, The Journal of biological chemistry.

[48]  L. Liotta,et al.  Tumor invasion and metastasis: an imbalance of positive and negative regulation. , 1991, Cancer research.

[49]  M. Duffy.  Urokinase Plasminogen Activator and Its Inhibitor, Pai-1, as Prognostic Markers in Breast Cancer: from Pilot to Level 1 Evidence Studies , 2002 .

[50]  C. Evans,et al.  MP24-07 NF-KAPPAB2/P52 INDUCES RESISTANCE TO ENZALUTAMIDE IN PROSTATE CANCER: ROLE OF ANDROGEN RECEPTOR AND ITS VARIANTS , 2014 .

[51]  Mei Han,et al.  Celecoxib and acetylbritannilactone interact synergistically to suppress breast cancer cell growth via COX-2-dependent and -independent mechanisms , 2011, Cell Death and Disease.