tranilast and Breast-Neoplasms

Hyaluronan is abundant in the extracellular matrix of many desmoplastic tumors and determines in large part the tumor biochemical and mechanical microenvironment. Additionally, it has been identified as one of the major physiological barriers to the effective delivery of drugs to solid tumors and its targeting with the use of pharmaceutical agents has shown to decompress tumor blood vessels, and thus improve tumor perfusion and efficacy of cytotoxic drugs. In this study, we investigated the contribution of hyaluronan to the accumulation of mechanical forces in tumors. Using experimental data from two orthotopic breast tumor models and treating tumors with two clinically approved anti-fibrotic drugs (tranilast and pirfenidone), we found that accumulation of growth-induced, residual forces in tumors are associated with hyaluronan content. Furthermore, mechanical characterization of the tumors revealed a good correlation of the accumulated forces with the elastic modulus of the tissue. Our results provide important insights on the mechano-pathology of solid tumors and can be used for the design of therapeutic strategies that target hyaluronan.

Topics: Adipose Tissue; Animals; Antineoplastic Agents; Breast Neoplasms; Cell Line, Tumor; Elastic Modulus; Female; Humans; Hyaluronic Acid; Mammary Glands, Animal; Mice; Mice, Inbred BALB C; Mice, Nude; Models, Biological; Neoplasm Transplantation; ortho-Aminobenzoates; Pyridones

This study was aimed at examining a separate or combined effect of tamoxifen and tranilast drugs on growth and proliferation of breast cancer cells.. Breast cancer is one of the most common cancers and the second leading cause of cancer death among women worldwide. Tamoxifen is the most widely used anti-estrogen for the treatment of breast cancer. Studies show that a combination therapy with other drugs enhances the activity of tamoxifen. Tranilast is an anti-inflammatory drug. We hypothesize that tranilast plus tamoxifen can work synergistically and help getting better result from this anticancer drug.. Two breast cancer cell lines, MCF-7 and MDA-MB-231, were treated with graduated concentrations of tamoxifen and tranilast alone or in combination at 24, 48 or 72 hours for MCF-7, and 48 hours for MDA-MB-231 cells. We used the MTT assay and lactate dehydrogenase leakage (LDH) assay to evaluate cell viability and cytotoxicity, respectively.. In both ER-positive and ER-negative breast cancer cell lines, the combination of tranilast and tamoxifen was more effective in growth inhibition than single drug exposure.. We have demonstrated that by means of a synergistic/additive inhibitory effect, tranilast was capable of enhancing the in vitro activity of tamoxifen on breast cancer cell lines. Based on the results obtained in this study, tranilast could be a candidate drug for combination therapy in resistant breast cancer patients (Fig. 9, Ref. 17).. breast cancer, Tamoxifen, Tranilast, LDH release, MTT.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Antineoplastic Agents, Hormonal; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Drug Synergism; Female; Humans; ortho-Aminobenzoates; Tamoxifen; Tumor Cells, Cultured

Tamoxifen is the most widely used anti-estrogen for the treatment of breast cancer. Studies show that the combination therapy with other substances that helps the activity of tamoxifen. The objective of this study was to evaluate the effect of tamoxifen when used in combination with tranilast on human breast cancer cells.. Two MCF-7 and MDA-MB-231 human breast cancer cell lines were treated with tamoxifen and/or tranilast. The cell viability and cytotoxicity was assessed using MTT and LDH assays; the apoptotic effects were examined by TUNEL assay, acridine orange/ethidium bromide staining and DNA laddering, also the expression levels of bax and bcl-2 genes were detected by real-time RT-PCR. The mRNA expression of TGF-β ligands and receptors examined using real-time RT-PCR and TGF-β1 protein secretion levels were also evaluated by ELISA assay. Inhibitory effect of these drugs on invasion and metastasis were tested by wound healing and matrigel invasion assay.. These findings indicate that tranilast, by synergistic effect, enhances the activity of tamoxifen and the TGF-β pathway is a target for this combination therapy, therefore; we propose that this combined treatment may be suitable selection in prevention of breast cancer.

Topics: Apoptosis; bcl-2-Associated X Protein; Breast Neoplasms; Cell Proliferation; Drug Synergism; Female; Gene Expression Regulation, Neoplastic; Humans; MCF-7 Cells; ortho-Aminobenzoates; Proto-Oncogene Proteins c-bcl-2; Tamoxifen; Transforming Growth Factor beta

Vascular endothelial growth factor and matrix metalloproteinases are two important factors for angiogenesis associated with breast cancer growth and progression. The present study was aimed to examine the effects of tamoxifen and tranilast drugs singly or in combination on proliferation of breast cancer cells and also to evaluate VEGF and MMP-9 expression and VEGF secretion levels.. Human breast cancer cell lines, MCF-7 and MDA-MB-231, were treated with tamoxifen and/or tranilast alone or in combination and percentage cell survival and proliferative activity were evaluated using LDH leakage and MTT assays. mRNA expression and protein levels were examined by real-time RT-PCR and ELISA assay, respectively.. LDH and MTT assays showed that the combined treatment of tamoxifen and tranilast resulted in a significant decrease in cell viability and cell proliferation compared with tamoxifen or tranilast treatment alone, with significant decrease in VEGF mRNA and protein levels. We also found that tamoxifen as a single agent rarely increased MMP-9 expression. A decrease in MMP-9 expression was seen after treatment with tranilast alone and in the combined treatment MMP-9 mRNA level was decreased.. This combination treatment can able to inhibit growth, proliferation and angiogenesis of breast cancer cells.

Topics: Anti-Inflammatory Agents, Non-Steroidal; Antineoplastic Agents, Hormonal; Apoptosis; Breast Neoplasms; Cell Proliferation; Drug Synergism; Enzyme-Linked Immunosorbent Assay; Female; Gene Expression Regulation, Neoplastic; Humans; L-Lactate Dehydrogenase; Matrix Metalloproteinase 9; ortho-Aminobenzoates; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tamoxifen; Tumor Cells, Cultured; Vascular Endothelial Growth Factor A

Cancer stem cells (CSCs) initiate tumors and have a high resistance to conventional cancer therapy. Tranilast is an orally active drug of low toxicity that exerts inhibitory effects on breast CSCs. This appears to depend on its aryl hydrocarbon receptor (AHR) agonistic activity, but this receptor has diverse functions and it is unclear how CSCs are inhibited. CSCs generate tumor spheres in low-adherence cultures, and we employed the mammosphere-forming assay as a functional test for breast CSCs. Because NF-κB has a key role in mammosphere formation and CSC-mediated tumor initiation, we examined that pathway. We also examined the role of neuropilin-1 (Nrp1), which is a growth factor coreceptor linked to the tumorigenicity of some CSCs. We found that tranilast concurrently suppressed mammosphere formation, Nrp1 expression and constitutive NF-κB activation. Flow cytometric analysis revealed that a subpopulation of breast cancer cells bearing breast CSC markers also expressed Nrp1. A blocking anti-Nrp1 antibody suppressed mammosphere formation. We examined whether there was a link between Nrp1 and NF-κB activation. The siRNA knockdown of Nrp1 severely suppressed NF-κB activation and mammosphere formation. The phosphorylation of Akt and ERK1/2 was also reduced, but to a lesser extent. We conclude that Nrp1 plays a key role in mammosphere formation and this activity is linked to NF-κB activation. Thus, Nrp1 might be a target for therapy against breast CSCs, and the anticancer drug tranilast suppresses its expression.

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Female; Gene Knockdown Techniques; Humans; Mice; Mitogen-Activated Protein Kinase 3; Neoplastic Stem Cells; Neuropilin-1; NF-kappa B; ortho-Aminobenzoates; Proto-Oncogene Proteins c-akt; RNA, Small Interfering; Spheroids, Cellular

In the treatment of breast cancer, although a wide of choice of drugs and treatment modalities are available, drug resistance or drug toxicity poses a considerable challenge. Tranilast is a well tolerated drug used in the treatment of allergic disorders. Previous works in various models have shown that tranilast has the potential to be used as an anti-cancer drug. Hence, in this study using human breast cancer cell lines BT-474 and MDA-MB-231, we studied the effect of tranilast on cell growth, migration and ability to prevent colony formation in vitro, properties that are relevant to a possible therapeutic effect in breast cancer. We found that tranilast inhibits the growth of both breast cancer cell lines. In the cell migration experiments, the tumor cells exhibit significantly slower wound closure after tranilast treatment, as well as reduced migration using an insert system. Downregulation of MRTF-A, a global cytoskeleton regulator was observed after tranilast treatment. Additionally, tranilast treatment increased levels of cleaved PARP in both cell lines tested indicating a stimulation of apoptosis. A significant reduction in colony size and number was observed in soft agar clonogenic assays in both cell lines after tranilast treatment. BT-474 cells were more responsive to tranilast treatment compared to MDA-MB-231 cells, suggesting a difference in modes of action, or sensitivity, possibly related to their different receptor status. Based on these changes in cancer cell lines, we conclude that tranilast exerts effects that set a rationale for future preclinical studies in animal models of breast cancer.

Topics: Antineoplastic Agents; Apoptosis; Breast Neoplasms; Cell Cycle; Cell Line, Tumor; Cell Movement; Cell Proliferation; Colony-Forming Units Assay; Dose-Response Relationship, Drug; Female; Humans; ortho-Aminobenzoates; Poly(ADP-ribose) Polymerases

Cancer stem cells (CSCs) have increased resistance to cancer chemotherapy. They can be enriched as drug-surviving CSCs (D-CSCs) by growth with chemotherapeutic drugs, and/or by sorting of cells expressing CSC markers such as aldehyde dehydrogenase-1 (ALDH). CSCs form colonies in agar, mammospheres in low-adherence cultures, and tumors following xenotransplantation in Scid mice. We hypothesized that tranilast, a non-toxic orally active drug with anti-cancer activities, would inhibit breast CSCs.. We examined breast cancer cell lines or D-CSCs generated by growth of these cells with mitoxantrone. Tranilast inhibited colony formation, mammosphere formation and stem cell marker expression. Mitoxantrone-selected cells were enriched for CSCs expressing stem cell markers ALDH, c-kit, Oct-4, and ABCG2, and efficient at forming mammospheres. Tranilast markedly inhibited mammosphere formation by D-CSCs and dissociated formed mammospheres, at pharmacologically relevant concentrations. It was effective against D-CSCs of both HER-2+ and triple-negative cell lines. Tranilast was also effective in vivo, since it prevented lung metastasis in mice injected i.v. with triple-negative (MDA-MB-231) mitoxantrone-selected cells. The molecular targets of tranilast in cancer have been unknown, but here we demonstrate it is an aryl hydrocarbon receptor (AHR) agonist and this plays a key role. AHR is a transcription factor activated by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), polycyclic aromatic hydrocarbons and other ligands. Tranilast induced translocation of the AHR to the nucleus and stimulated CYP1A1 expression (a marker of AHR activation). It inhibited binding of the AHR to CDK4, which has been linked to cell-cycle arrest. D-CSCs expressed higher levels of the AHR than other cells. Knockdown of the AHR with siRNA, or blockade with an AHR antagonist, entirely abrogated the anti-proliferative and anti-mammosphere activity of tranilast. Thus, the anti-cancer effects of tranilast are AHR dependent.. We show that tranilast is an AHR agonist with inhibitory effects on breast CSCs. It is effective against CSCs of triple-negative breast cancer cells selected for anti-cancer drug resistance. These results suggest it might find applications in the treatment of breast cancer.

Topics: Aldehyde Dehydrogenase; Animals; Anti-Inflammatory Agents, Non-Steroidal; Blotting, Western; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Cytochrome P-450 CYP1A1; Dose-Response Relationship, Drug; Female; Flow Cytometry; Humans; Lung Neoplasms; Mammary Neoplasms, Experimental; Mice; Mice, Inbred NOD; Mice, SCID; Neoplastic Stem Cells; Octamer Transcription Factor-3; ortho-Aminobenzoates; Receptors, Aryl Hydrocarbon; RNA Interference; Transplantation, Heterologous

In MCF-7 breast cancer cells, insulin-like growth factor-1 (IGF-1) increased the calcium-permeability of the cells by activating a voltage-independent calcium-permeable channel. IGF-1 also induced oscillatory elevation of cytoplasmic free calcium concentration in these cells. An anti-allergic compound, tranilast, reduced the calcium-permeability augmented by IGF-1 in a dose-dependent manner and blocked the oscillatory elevation of cytoplasmic free calcium concentration. Tranilast did not affect early intracellular signals activated by IGF-1, including receptor autophosphorylation, activations of Ras, mitogen-activated protein kinase and phosphatidylinositol 3-kinase. Tranilast inhibited increases in [3H]-thymidine incorporation, DNA content and cell number induced by IGF-1. The ID50 for [3H]-thymidine incorporation and DNA content were about 10 microM. The inhibitory effect of tranilast was reversible, and cell viability was not affected. Treatment with tranilast increased the number of cells in the G1 phase suggesting that this compound induced G1 arrest. Tranilast also reduced the phosphorylation of the retinoblastoma protein. These results indicate that tranilast inhibits the IGF-1-induced cell growth in MCF-7 cells by blocking calcium entry.

Topics: Breast Neoplasms; Calcium; Calcium Channel Blockers; Cell Division; Cell Survival; DNA Fragmentation; DNA Replication; DNA, Neoplasm; Humans; Insulin-Like Growth Factor I; ortho-Aminobenzoates; Patch-Clamp Techniques; Phosphorylation; Retinoblastoma Protein; Tumor Cells, Cultured