4-(5-benzo(1-3)dioxol-5-yl-4-pyridin-2-yl-1h-imidazol-2-yl)benzamide and Neoplasm-Metastasis

Accumulating evidences have demonstrated that mesenchymal stem cells (MSC) could be recruited to the tumor microenvironment. Umbilical cord mesenchymal stem cells (UCMSC) were attractive vehicles for delivering therapeutic agents against cancer. Nevertheless, the safety of UCMSC in the treatment of tumors including hepatocellular carcinoma (HCC) was still undetermined.. In this study, an in vitro co-culture system was established to evaluate the effect of UCMSC on the cell growth, cancer stem cell (CSC) characteristics, drug resistance, metastasis of 3D-cultured HCC cells, and the underlying mechanism was also investigated.. It was found that after co-cultured with UCMSC, the metastatic ability of 3D-cultured HCC cells was significantly enhanced as indicated by up-regulation of matrix metalloproteinase (MMP), epithelial-mesenchymal transition (EMT)-related genes, and migration ability. However, cell growth, drug resistance and CSC-related gene expression of HCC cells were not affected by UCMSC. Moreover, EMT was reversed, MMP-2 expression was down-regulated, and migration ability of HCC cell was significantly inhibited when TGF-β receptor inhibitor SB431542 was added into the co-culture system.. Therefore, these data indicated that UCMSC could significantly enhance the tumor cell metastasis, which was due to the EMT of HCC cells induced by TGF-β.

Topics: Benzamides; Carcinoma, Hepatocellular; Cell Line, Tumor; Cells, Cultured; Cisplatin; Coculture Techniques; Dioxoles; Drug Resistance, Neoplasm; Gene Expression Regulation, Neoplastic; Humans; Liver Neoplasms; Matrix Metalloproteinases; Mesenchymal Stem Cells; Neoplasm Metastasis; Transforming Growth Factor beta; Tumor Microenvironment

Cancer progression towards metastasis follows a defined sequence of events described as the metastatic cascade. For extravasation and transendothelial migration metastatic cells interact first with endothelial cells. Yet the role of endothelial cells during the process of metastasis formation and extravasation is still unclear, and the interaction between metastatic and endothelial cells during transendothelial migration is poorly understood. Since tumor cells are well known to express TGF-β, and the compact endothelial layer undergoes a series of changes during metastatic extravasation (cell contact disruption, cytoskeletal reorganization, enhanced contractility), we hypothesized that an EndMT may be necessary for metastatic extravasation. We demonstrate that primary cultured rat brain endothelial cells (BEC) undergo EndMT upon TGF-β1 treatment, characterized by the loss of tight and adherens junction proteins, expression of fibronectin, β1-integrin, calponin and α-smooth muscle actin (SMA). B16/F10 cell line conditioned and activated medium (ACM) had similar effects: claudin-5 down-regulation, fibronectin and SMA expression. Inhibition of TGF-β signaling during B16/F10 ACM stimulation using SB-431542 maintained claudin-5 levels and mitigated fibronectin and SMA expression. B16/F10 ACM stimulation of BECs led to phosphorylation of Smad2 and Smad3. SB-431542 prevented SMA up-regulation upon stimulation of BECs with A2058, MCF-7 and MDA-MB231 ACM as well. Moreover, B16/F10 ACM caused a reduction in transendothelial electrical resistance, enhanced the number of melanoma cells adhering to and transmigrating through the endothelial layer, in a TGF-β-dependent manner. These effects were not confined to BECs: HUVECs showed TGF-β-dependent SMA expression when stimulated with breast cancer cell line ACM. Our results indicate that an EndMT may be necessary for metastatic transendothelial migration, and this transition may be one of the potential mechanisms occurring during the complex phenomenon known as metastatic extravasation.

Topics: Animals; Benzamides; Brain; Cell Line, Tumor; Cells, Cultured; Culture Media, Conditioned; Dioxoles; Endothelial Cells; Epithelial-Mesenchymal Transition; Gene Expression Regulation; Human Umbilical Vein Endothelial Cells; Humans; MCF-7 Cells; Mice; Neoplasm Metastasis; Rats; Transforming Growth Factor beta1

The transforming growth factor beta (TGFβ) signaling cascade is considered as one of the pivotal oncogenic pathways in most advanced cancers. Inhibition of the TGFβ signaling pathway by specific antagonists, neutralizing antibodies, or small molecules is considered as an effective strategy for the treatment of tumor growth and metastasis. Here we demonstrated the identification of a series of tetrahydro-β-carboline derivatives from virtual screening which potentially inhibit the TGFβ signaling pathway. Optimization of the initial hit compound 2-benzoyl-1,3,4,9-tetrahydro-β-carboline (8a) through substitution at different positions to define the structure-activity relationship resulted in the discovery of potent inhibitors of the TGFβ signaling pathway. Among them, compound 8d, one of the tested compounds, not only showed potent inhibition of lung cancer cell proliferation and migration in vitro but also strongly suppressed growth of lung cancer and breast cancer in vivo.

Topics: Animals; Antineoplastic Agents; Carbolines; Cell Line, Tumor; Cell Movement; Cell Proliferation; Drug Screening Assays, Antitumor; Female; Humans; Lung Neoplasms; Mammary Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Mice, Nude; Molecular Docking Simulation; Neoplasm Metastasis; Neoplasm Transplantation; Signal Transduction; Structure-Activity Relationship; Transforming Growth Factor beta

To examine whether ionizing radiation enhances the migratory and invasive abilities of cancer cells through transforming growth factor (TGF-β)-mediated epithelial-mesenchymal transition (EMT).. Six cancer cell lines originating from different human organs were irradiated by 60Co γ-ray at a total dose of 2 Gy, and the changes associated with EMT, including morphology, EMT markers, migration and invasion, were observed by microscope, Western blot, immunofluorescence, scratch assay, and transwell chamber assay, respectively. Then the protein levels of TGF-β in these cancer cells were detected by enzyme-linked immunosorbent assay, and the role of TGF-β signaling pathway in the effect of ionizing radiation on EMT was investigate by using the specific inhibitor SB431542.. After irradiation with γ-ray at a total dose of 2 Gy, cancer cells presented the mesenchymal phenotype, and compared with the sham-irradiation group the expression of epithelial markers was decreased and of mesenchymal markers was increased, the migratory and invasive capabilities were strengthened, and the protein levels of TGF-β were enhanced. Furthermore, events associated with EMT induced by IR in A549 could be reversed through inhibition of TGF-β signaling.. These results suggest that EMT mediated by TGF-β plays a critical role in IR-induced enhancing of migratory and invasive capabilities in cancer cells.

Topics: Benzamides; Cell Line, Tumor; Cell Movement; Cobalt Radioisotopes; Dioxoles; Epithelial-Mesenchymal Transition; Gamma Rays; Humans; Neoplasm Invasiveness; Neoplasm Metastasis; Receptors, Transforming Growth Factor beta; Transforming Growth Factor beta1

Human peritoneal mesothelial cells (HPMCs) in intact mesothelium have been demonstrated to protect against tumor peritoneal metastasis. We have previously reported that gastric cancer cells can induce peritoneal apoptosis, lead to damage of peritoneum integrity, and therefore promote peritoneal metastasis. In this study, we investigated the effects of TGF-beta1 on tumor-mesothelial interaction. Briefly, the levels of various soluble factors, in particular TGF-beta1, were measured. HMrSV5 cells, a human peritoneal mesothelial cell line, were co-incubated with TGF-beta1, gastric cancer cells, or gastric cancer cells and TGF-beta1 receptor inhibitor SB431542. The expressions of smad 2/3 and phosphorylated smad 2/3, indicator of TGF-beta/Smads pathway activation, were evaluated. Then the morphological changes of HPMCs were observed. The cell damage was quantitatively determined by fluorescent microscopy and flow cytometry. Tumor-mesothelial cell adhesion was also examined. Results showed a significant elevation of TGF-beta1 expression, which is companied by dramatically increased phosphorylated-smad 2/3 levels, after mesothelial cell co-culture with the gastric cancer cell line. In addition, mesothelial cells exposed to gastric cancer cells or TGF-beta1 became exfoliated and exhibited signs of injury, while blocking TGF-beta1 can partially inhibit these effects. These results indicate that soluble factors, such as TGF-beta1, produced in autocrine/paracrine manner in the peritoneal cavity, affect the morphology and function of mesothelial cells so that the resulting environment becomes favorable for peritoneal metastases.

Topics: Apoptosis; Autocrine Communication; Benzamides; Carcinoma; Cell Adhesion; Cell Shape; Cells, Cultured; Coculture Techniques; Culture Media, Conditioned; Culture Media, Serum-Free; Dioxoles; Epithelial Cells; Epithelium; Humans; Neoplasm Metastasis; Paracrine Communication; Peritoneal Neoplasms; Receptors, Transforming Growth Factor beta; Stomach Neoplasms; Transforming Growth Factor beta1

Immunosuppressed individuals undergoing organ transplantation are at increased risk of recurrences of initial cancers, although how immunosuppressive therapy increases early cancer metastasis remains unclear.. The metastatic fate of luciferase-expressing rat metastatic colon cancer cells (luc-RCN-H4) injected intravenously into the liver of syngeneic and allogeneic rats was examined in the presence of the immunosuppressant cyclosporin A (CsA) by in vivo luminescent technique. With respect to potential tumor-progressing factors, contribution of chemokine receptors and transforming growth factor (TGF)-beta1 to early metastasis was evaluated using their specific signaling inhibitors.. F344 rats injected in the liver with luc-RCN-H4 cells did not always exhibit the formation of tumors and showed a dormant state as long as 60 days after inoculation without CsA. However, CsA released early luc-RCN-H4 cells from dormancy within 2 weeks at nearly 100% in liver and preferentially promoted metastasis to the lymph nodes (approximately 40%). A similar dissemination occurred even in minor histocompatibility complex-disparate hosts. As a tumor-progressing factor, RCN-H4 cells aberrantly expressed chemokine receptors CXCR4 and CCR7. The chemokine receptor (CXC) R4-specific antagonist AMD3100 decreased early metastasis of luc-RCN-H4 cells in rats with ischemic liver conditions (P<0.05), but CsA treatment did not enhance early adhesion. Use of CsA was able to facilitate TGF-beta1 expression and the subsequent TGF-beta-mediated random migration was blocked by the use of the specific signaling inhibitor SB431542 in vitro.. Whereas the chemokine receptor expression by cancer cells is implicated with early organotropic dissemination even under CsA-mediated immune suppression, rather, CsA enhances the late-phase progression after tumor adhesion through TGF-beta1 expression.

Topics: Adenocarcinoma; Animals; Benzamides; Blotting, Western; Cell Adhesion; Cell Line, Tumor; Cell Movement; Colonic Neoplasms; Cyclosporine; Dioxoles; Disease Progression; Gene Expression Regulation, Neoplastic; Image Processing, Computer-Assisted; Killer Cells, Natural; Liver Neoplasms; Luminescence; Lymphatic Metastasis; Male; Neoplasm Metastasis; Rats; Rats, Inbred F344; Receptors, Chemokine; Reperfusion Injury; Reverse Transcriptase Polymerase Chain Reaction; Transforming Growth Factor beta; Transforming Growth Factor beta1

Small molecule inhibitors of signaling pathways have proven to be extremely useful for the development of therapeutic strategies for human cancers. Blocking the tumor-promoting effects of transforming growth factor-beta (TGF-beta) in advanced stage carcinogenesis provides a potentially interesting drug target for therapeutic intervention. Although very few TGF-beta receptor kinase inhibitors (TRKI) are now emerging in preclinical studies, nothing is known about how these inhibitors might regulate the tumor-suppressive or tumor-promoting effects of TGF-beta, or when these inhibitors might be useful for treatment during cancer progression. We have investigated the potential of TRKI in new therapeutic approaches in preclinical models. Here, we demonstrate that the TRKI, SB-431542, inhibits TGF-beta-induced transcription, gene expression, apoptosis, and growth suppression. We have observed that SB-431542 attenuates the tumor-promoting effects of TGF-beta, including TGF-beta-induced EMT, cell motility, migration and invasion, and vascular endothelial growth factor secretion in human cancer cell lines. Interestingly, SB-431542 induces anchorage independent growth of cells that are growth-inhibited by TGF-beta, whereas it reduces colony formation by cells that are growth-promoted by TGF-beta. However, SB-431542 has no effect on a cell line that failed to respond to TGF-beta. This represents a novel potential application of these inhibitors as therapeutic agents for human cancers with the goal of blocking tumor invasion, angiogenesis, and metastasis, when tumors are refractory to TGF-beta-induced tumor-suppressor functions but responsive to tumor-promoting effects of TGF-beta.

Topics: Activin Receptors, Type I; Animals; Antineoplastic Agents; Apoptosis; Benzamides; Blotting, Western; Cell Adhesion; Cell Line; Cell Line, Tumor; Cell Movement; Dioxoles; Enzyme Inhibitors; Gene Expression Regulation, Neoplastic; Humans; Immunoprecipitation; Mice; Mink; Models, Biological; Neoplasm Invasiveness; Neoplasm Metastasis; Neovascularization, Pathologic; Protein Serine-Threonine Kinases; Rats; Receptor, Transforming Growth Factor-beta Type I; Receptors, Transforming Growth Factor beta; Time Factors; Transcription, Genetic; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A