Phenethyl isothiocyanate inhibits migration and invasion of human gastric cancer AGS cells through suppressing MAPK and NF-kappaB signal pathways.

Cell motility involves metastasis suppressors and other regulators that play an important role in tumor invasion and metastasis. Phenethyl isothiocyanate (PEITC), found in dietary cruciferous vegetables, has been found to exhibit antitumor properties and therefore is of special interest for the development of chemopreventive and chemotherapeutic agent for human cancers. Here, we report that in addition to its function as an anticancer agent, and PEITC can inhibit migration and invasion through the extracellular signal-regulated kinases 1/2 (ERK1/2), protein kinase C (PKC) and nuclear factor-kappaB (NF-kappaB) signaling pathways in human gastric cells. The results from wound healing and Boyden chamber assays (migration and invasion) assay indicated that PEITC exhibited an inhibitory effect on the migration and invasion of AGS cells. Results from Western blotting examination demonstrated that PEITC exerted an inhibitory effect on the ERK1/2, mitogen-activated protein kinase kinase 7 (MKK7), MAP kinase kinase kinase 3 (MEKK3), son of sevenless 1 (SOS1), PKC, Ras homolog gene family, member A (Rho A) and urokinase-type plasminogen activator (uPA), causing the inhibition of matrix metallopeptidase-2 (MMP-2) and -9 then followed by the inhibition of invasion and migration of GAS cells in vitro. PEITC also inhibited Ras, growth factor receptor-bound protein 2 (GRB2), vascular endothelial growth factor (VEGF), focal adhesion kinase (FAK), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), causing inhibition of cell proliferation of AGS cells. Results from real-time PCR showed that PEITC inhibited the gene expressions of MMP-2, -7 and -9, FAK and RhoA after PEITC treatment for 24 and 48 h of AGS cells. Taken together, these findings may provide insight into a new mechanisms and functions of PEITC in migration and invasion of human gastric cancer AGS cells. Our data imply that molecular targeting of PKC leading to the inhibition of MMP-2 and -9 might be a useful strategy for the inhibition of migration and invasion of human gastric cancer.

[1]  J. Hayes,et al.  Dietary indoles and isothiocyanates that are generated from cruciferous vegetables can both stimulate apoptosis and confer protection against DNA damage in human colon cell lines. , 2001, Cancer research.

[2]  Ji Won Lee,et al.  Phenethyl isothiocyanate induced apoptosis via down regulation of Bcl-2/XIAP and triggering of the mitochondrial pathway in MCF-7 cells , 2008, Archives of pharmacal research.

[3]  J. Fuselier,et al.  A conjugate of camptothecin and a somatostatin analog against prostate cancer cell invasion via a possible signaling pathway involving PI3K/Akt, αVβ3/αVβ5 and MMP-2/-9 , 2007 .

[4]  V. Mersch-Sundermann,et al.  Induction of apoptosis in tumor cells by naturally occurring sulfur-containing compounds. , 2005, Mutation research.

[5]  S. Amin,et al.  Effects of aromatic isothiocyanates on tumorigenicity, O6-methylguanine formation, and metabolism of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone in A/J mouse lung. , 1989, Cancer research.

[6]  Yi Tang,et al.  Extracellular matrix metalloproteinase inducer (EMMPRIN) and matrix metalloproteinases (MMPs) as regulators of tumor–host interaction in a spontaneous metastasis model in rats , 2008, Histochemistry and Cell Biology.

[7]  T. Kensler,et al.  Anticarcinogenic activities of sulforaphane and structurally related synthetic norbornyl isothiocyanates. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[8]  T. Hsia,et al.  DNA damage and endoplasmic reticulum stress mediated curcumin-induced cell cycle arrest and apoptosis in human lung carcinoma A-549 cells through the activation caspases cascade- and mitochondrial-dependent pathway. , 2008, Cancer letters.

[9]  K. Shirouzu,et al.  The significance of MMP-1 and MMP-2 in peritoneal disseminated metastasis of gastric cancer , 2010, Surgery Today.

[10]  J. Ajani.  Evolving chemotherapy for advanced gastric cancer. , 2005, The oncologist.

[11]  Jai-Sing Yang,et al.  Curcumin blocks migration and invasion of mouse-rat hybrid retina ganglion cells (N18) through the inhibition of MMP-2, -9, FAK, Rho A and Rock-1 gene expression. , 2010, Oncology reports.

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

[13]  R. Kalluri,et al.  Renal fibrosis. Extracellular matrix microenvironment regulates migratory behavior of activated tubular epithelial cells. , 2002, The American journal of pathology.

[14]  J. Pullar,et al.  Phenethyl isothiocyanate triggers apoptosis in Jurkat cells made resistant by the overexpression of Bcl-2. , 2006, Cancer research.

[15]  M. Walz,et al.  Matrix metalloproteinase inhibitor RO 28-2653 decreases liver metastasis by reduction of MMP-2 and MMP-9 concentration in BOP-induced ductal pancreatic cancer in Syrian Hamsters: inhibition of matrix metalloproteinases in pancreatic cancer. , 2006, Prostaglandins, leukotrienes, and essential fatty acids.

[16]  A. Hölscher,et al.  Expression of MMP‐2 is associated with progression and lymph node metastasis of gastric carcinoma , 2001, Histopathology.

[17]  Kuang-Chi Lai,et al.  Benzyl isothiocyanate (BITC) inhibits migration and invasion of human colon cancer HT29 cells by inhibiting matrix metalloproteinase-2/-9 and urokinase plasminogen (uPA) through PKC and MAPK signaling pathway. , 2010, Journal of agricultural and food chemistry.

[18]  K. Sugimachi,et al.  Matrix metalloproteinase-7 expression in gastric carcinoma. , 1996, Gut.

[19]  M. Fan,et al.  Emodin induces apoptosis of human tongue squamous cancer SCC-4 cells through reactive oxygen species and mitochondria-dependent pathways. , 2009, Anticancer research.

[20]  O. Leoni,et al.  In vitro antiproliferative activity of isothiocyanates and nitriles generated by myrosinase-mediated hydrolysis of glucosinolates from seeds of cruciferous vegetables. , 2000, Journal of agricultural and food chemistry.

[21]  S. Chu,et al.  Silibinin inhibits cell invasion through inactivation of both PI3K-Akt and MAPK signaling pathways. , 2005, Chemico-biological interactions.

[22]  H. Kwaan.  The plasminogen-plasmin system in malignancy , 1992, Cancer and Metastasis Reviews.

[23]  S. Gruber,et al.  Direct evidence linking expression of matrix metalloproteinase 9 (92-kDa gelatinase/collagenase) to the metastatic phenotype in transformed rat embryo cells. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Kuang-Chi Lai,et al.  Berberine suppresses in vitro migration and invasion of human SCC-4 tongue squamous cancer cells through the inhibitions of FAK, IKK, NF-kappaB, u-PA and MMP-2 and -9. , 2009, Cancer letters.

[25]  M. Macías-Silva,et al.  Src kinase regulates metalloproteinase-9 secretion induced by type IV collagen in MCF-7 human breast cancer cells. , 2008, Matrix biology : journal of the International Society for Matrix Biology.

[26]  N. Swamy,et al.  Phenethyl isothiocyanate (PEITC) inhibits growth of ovarian cancer cells by inducing apoptosis: role of caspase and MAPK activation. , 2006, Gynecologic oncology.

[27]  Y. Seyama,et al.  Matrix metalloproteinase-2 and -9 in bile as a marker of liver metastasis in colorectal cancer. , 2001, Biochemical and biophysical research communications.

[28]  K. Brown,et al.  Induction of apoptosis by phenethyl isothiocyanate in cells overexpressing Bcl-XL. , 2008, Cancer letters.

[29]  M. Kurdoğlu,et al.  The relation of gelatinase (MMP-2 and -9) expression with distant site metastasis and tumour aggressiveness in colorectal cancer , 1999, British Journal of Cancer.

[30]  Young-Chae Chang,et al.  Ascofuranone suppresses PMA-mediated matrix metalloproteinase-9 gene activation through the Ras/Raf/MEK/ERK- and Ap1-dependent mechanisms. , 2006, Carcinogenesis.

[31]  Cheorl-Ho Kim,et al.  Suppression of PMA-induced tumor cell invasion by capillarisin via the inhibition of NF-kappaB-dependent MMP-9 expression. , 2008, Biochemical and biophysical research communications.

[32]  G. Kuttan,et al.  Amentoflavone Inhibits Experimental Tumor Metastasis Through a Regulatory Mechanism Involving MMP-2, MMP-9, Prolyl Hydroxylase, Lysyl Oxidase, VEGF, ERK-1, ERK-2, STAT-1, nm23 and Cytokines in Lung Tissues of C57BL/6 Mice , 2008, Immunopharmacology and immunotoxicology.

[33]  T. Koga,et al.  Clinicopathological features and overexpression of matrix metalloproteinases in intramucosal gastric carcinoma with lymph node metastasis. , 2000, Clinical cancer research : an official journal of the American Association for Cancer Research.

[34]  Yuesheng Zhang.  Cancer-preventive isothiocyanates: measurement of human exposure and mechanism of action. , 2004, Mutation research.

[35]  Po-Yuan Chen,et al.  Rhein induced apoptosis through the endoplasmic reticulum stress, caspase- and mitochondria-dependent pathways in SCC-4 human tongue squamous cancer cells. , 2009, In vivo.

[36]  M. Duffy,et al.  Urokinase plasminogen activator: A prognostic marker in multiple types of cancer , 1999, Journal of surgical oncology.

[37]  Y. Okada,et al.  Expression and tissue localization of matrix metalloproteinase 7 (matrilysin) in human gastric carcinomas. Implications for vessel invasion and metastasis , 1998, International journal of cancer.

[38]  H. Klocker,et al.  Type IV collagenase (matrix metalloproteinase-2 and -9) in prostate cancer , 2004, Prostate Cancer and Prostatic Diseases.

[39]  Wun-Jae Kim,et al.  Signaling pathway for TNF-alpha-induced MMP-9 expression: mediation through p38 MAP kinase, and inhibition by anti-cancer molecule magnolol in human urinary bladder cancer 5637 cells. , 2008, International immunopharmacology.

[40]  T. Wobbes,et al.  Matrix metalloproteinase 2 and 9 activity in patients with colorectal cancer liver metastasis , 2003, The British journal of surgery.

[41]  Larine Nee,et al.  TNF-Alpha and IL-1Beta-Mediated Regulation of MMP-9 and TIMP-1 in Human Glomerular Mesangial Cells , 2007, Nephron Experimental Nephrology.

[42]  Young-ho Kim,et al.  Resveratrol inhibits phorbol myristate acetate-induced matrix metalloproteinase-9 expression by inhibiting JNK and PKC δ signal transduction , 2004, Oncogene.

[43]  P. Plinkert,et al.  The p38 SAPK pathway regulates the expression of the MMP-9 collagenase via AP-1-dependent promoter activation. , 2001, Experimental cell research.

[44]  Jai-Sing Yang,et al.  Danthron inhibits the migration and invasion of human brain glioblastoma multiforme cells through the inhibition of mRNA expression of focal adhesion kinase, Rho kinases-1 and metalloproteinase-9. , 2009, Oncology reports.

[45]  J. Nelson,et al.  Caspase-Dependent Apoptosis Induction by Phenethyl Isothiocyanate, a Cruciferous Vegetable-Derived Cancer Chemopreventive Agent, Is Mediated by Bak and Bax , 2005, Clinical Cancer Research.

[46]  S. Chakraborti,et al.  Regulation of matrix metalloproteinases: An overview , 2003, Molecular and Cellular Biochemistry.

[47]  G. Murray,et al.  Matrix metalloproteinases and their inhibitors in gastric cancer , 1998, Gut.