Benzyl-isothiocyanate (BITC) is a hydrolysis product of glucotropaeolin, a compound found in cruciferous vegetables, and has also been shown to have anti-tumor properties. To evaluate the effects of BITC administration on the tumor growth and metastasis of breast cancer, 4T1 murine mammary carcinoma cells were injected into the inguinal mammary fat pads of syngeneic female BALB/c mice. One day later, the mice were subjected to gavage for 4 weeks with BITC (0, 5, or 10 mg/kg body weight/day). Oral BITC treatment induced a significant reduction in the growth of solid tumors. BITC reduced hemoglobin contents and CD31 and vascular endothelial growth factor (VEGF) expression in the tumors, as well as circulating levels of VEGF. Reduced expressions of proliferating cell nuclear antigen and cyclin-dependent kinase 4 were noted in the tumors of BITC-treated mice. BITC markedly increased the numbers of apoptotic cells with increased Bax expression, cleaved caspase-3, and PARP levels, but reduced Bcl-2 expression in tumor tissues. In addition, BITC was shown to reduce the numbers of pulmonary tumor nodules and the total pulmonary metastatic volume. BITC induced a significant reduction in the levels of matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of metalloproteinase (TIMP)-1, and urokinase-type plasminogen activator in the sera and lungs of 4T1 cell-injected mice. However, the concentrations of TIMP-2 and plasminogen activator inhibitor-1 were increased in the sera and lungs of BITC-treated mice. The results of this study indicate that BITC has potential as a preventive agent for metastatic breast cancer.

Benzyl isothiocyanate (BITC), which is found in cruciferous vegetables, has been shown to have anti-carcinogenic properties. Hepatocyte growth factor (HGF) has the ability to stimulate dissociation, migration, and invasion in various tumor cells, and abnormally increased expressions of HGF and its transmembrane tyrosine kinase receptor, c-Met, have previously been detected in human breast cancer, and are associated with high tumor grade and poor prognosis. In this study, in order to assess the mechanisms relevant to the BITC-induced regulation of breast cancer cell migration and invasion, MDA-MB-231 human breast cancer cells and 4T1 murine mammary carcinoma cells were cultured in the presence of 0-4 μmol/l BITC with or without 10 μg/l of HGF. BITC inhibited both the basal and HGF-induced migration of MDA-MB-231 and 4T1 cells in a dose-dependent manner. In MDA-MB-231 cells, BITC reduced both basal and HGF-induced secretion and activity of urokinase-type plasminogen activator (uPA). In addition, BITC increased the protein levels of plasminogen activator inhibitor-1. HGF stimulated c-Met and Akt phosphorylation, but did not affect the phosphorylation of extracellular signal-regulated kinase-1/2 or stress-activated protein/c-jun N-terminal kinase. BITC suppressed NF-κB activity and reduced the HGF-induced phosphorylation of c-Met and Akt in a dose-dependent manner. LY294002, a specific Akt inhibitor, reduced both basal and HGF-induced uPA secretion and migration of MDA-MB-231 cells. In this study, we demonstrated that BITC profoundly inhibits the migration and invasion of MDA-MB-231 cells, which is associated with reduced uPA activity, and also that these phenomena are accompanied by the suppression of Akt signaling.

Benzyl isothiocyanate (BITC) is a hydrolysis product of glucotropaeolin found in cruciferous vegetables, and has been reported to have anti-tumor properties. In our previous study, we observed that BITC inhibited growth and metastatic capacity of breast cancer cells in vitro cell culture models. The present study examined the in vivo effects of BITC administration on tumor growth and metastasis of breast cancer cells. To establish the in vivo breast cancer model, 4T1 mammary carcinoma cells (5 × 104 cells) were injected into the inguinal mammary fat pad of syngeneic female BALB/c mice. One day later, the mice were divided into 3 groups and subjected to gavage with BITC (0, 5 or 10 mg/kg body weight/day) for 4 weeks. Oral administration of BITC significantly reduced the volume and weight of solid 4T1 cell tumors in BALB/c mice. BITC administration decreased the expression of proliferating cell nuclear antigen, Bcl-2, CD31 and VEGF, but increased the levels of Bax, cleaved caspase-3, −7, −9 and cleaved PARP in the tumors. In addition, Oral administration of BITC resulted in a significant reduction in the numbers of pulmonary tumor nodules and total pulmonary metastatic volume. To understand the mechanisms by which BITC exerts antimetastatic effects, metastasis-related proteins in the sera and lungs were measured by utilizing the relevant ELISA kits and Western blot analysis, respectively. BITC treatment significantly decreased the concentrations of matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of metalloproteinase (TIMP)-1, and urokinase-type plasminogen activator (uPA) in the sera of mice injected with 4T1 cells. However, the concentrations of TIMP-2 and plasminogen activator inhibitor (PAI)-1 were increased in the sera of mice treated with BITC. Additionally, the administration of BITC reduced the levels of MMP-2, MMP-9, TIMP-1, and uPA and increased TIMP-2 and PAI-1 in the lungs of these animals. In this study, we have demonstrated that BITC profoundly suppresses the tumor growth and lung metastasis in mice bearing orthotopic xenografts of 4T1 breast cancer cells. The present results indicate that BITC has potential as an anti-cancer agent for the treatment of breast cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1444.