ABSTRACT
The aim of the present study was to verify the effects of regorafenib on apoptosis and metastatic potential in TSGH 8301 human bladder carcinoma cells in vitro. Cells were treated with different concentration of regorafenib for different periods of time. Effects of regorafenib on cell viability, apoptosis pathways, metastatic potential, and expression of metastatic and anti-apoptotic proteins were evaluated with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay, flow cytometry, cell migration and invasion assay, and western blotting. We found regorafenib significantly reduced cell viability, cell migration and invasion, and expression of metastatic and anti-apoptotic proteins. In addition, regorafenib significantly induced accumulation of sub-G1 phase cells, loss of mitochondrial membrane potential, and expression of active caspase-3 and caspase-8. These results show that regorafenib not only induces apoptosis, but also inhibits metastatic potential in bladder cancer TSGH 8301 cells in vitro.
Subject(s)
Apoptosis/drug effects , Cell Movement/drug effects , Phenylurea Compounds/pharmacology , Pyridines/pharmacology , Urinary Bladder Neoplasms/pathology , Cell Proliferation/drug effects , Humans , Membrane Potential, Mitochondrial/drug effects , Tumor Cells, Cultured , Urinary Bladder Neoplasms/drug therapy , Urinary Bladder Neoplasms/metabolismABSTRACT
Acute hepatic failure (AHF) is a severe liver injury leading to sustained damage and complications. Induced pluripotent stem cells (iPSCs) may be an alternative option for the treatment of AHF. In this study, we reprogrammed human dental pulp-derived fibroblasts into iPSCs, which exhibited pluripotency and the capacity to differentiate into tridermal lineages, including hepatocyte-like cells (iPSC-Heps). These iPSC-Heps resembled human embryonic stem cell-derived hepatocyte-like cells in gene signature and hepatic markers/functions. To improve iPSC-Heps engraftment, we next developed an injectable carboxymethyl-hexanoyl chitosan hydrogel (CHC) with sustained hepatocyte growth factor (HGF) release (HGF-CHC) and investigated the hepatoprotective activity of HGF-CHC-delivered iPSC-Heps in vitro and in an immunocompromised AHF mouse model induced by thioacetamide (TAA). Intrahepatic delivery of HGF-CHC-iPSC-Heps reduced the TAA-induced hepatic necrotic area and rescued liver function and recipient viability. Compared with PBS-delivered iPSC-Heps, the HGF-CHC-delivered iPSC-Heps exhibited higher antioxidant and antiapoptotic activities that reduced hepatic necrotic area. Importantly, these HGF-CHC-mediated responses could be abolished by administering anti-HGF neutralizing antibodies. In conclusion, our findings demonstrated that HGF mediated the enhancement of iPSC-Hep antioxidant/antiapoptotic capacities and hepatoprotection and that HGF-CHC is as an excellent vehicle for iPSC-Hep engraftment in iPSC-based therapy against AHF.
Subject(s)
Cell Differentiation/drug effects , Hepatocyte Growth Factor/pharmacology , Hepatocytes/cytology , Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Induced Pluripotent Stem Cells/transplantation , Liver Failure, Acute/therapy , Liver Regeneration , Alanine Transaminase/analysis , Animals , Antioxidants/chemistry , Antioxidants/metabolism , Aspartate Aminotransferases/analysis , Bilirubin/analysis , Cells, Cultured , Cellular Reprogramming , Chitosan/analogs & derivatives , Chitosan/chemistry , Dental Pulp/cytology , Female , Hepatocyte Growth Factor/chemistry , Hepatocyte Growth Factor/metabolism , Hepatocytes/metabolism , Humans , Induced Pluripotent Stem Cells/cytology , Liver/metabolism , Liver Failure, Acute/chemically induced , Liver Failure, Acute/pathology , Male , Malondialdehyde , Mice , Mice, Inbred BALB C , Mice, Inbred ICR , Mice, Nude , Reactive Oxygen Species/metabolism , Thioacetamide/toxicityABSTRACT
Induced pluripotent stem cells (iPSCs) are generated by reprogramming somatic cells to a pluripotent state by the introduction of specific factors. They can be generated from cells of different origins, such as fibroblasts, keratinocytes, hepatocytes, and blood. iPSCs are similar to embryonic stem cells (ESCs) in several aspects, such as morphology, expression of pluripotency markers, and the ability to develop teratoma that contains tissue from three germ layers. In addition, iPSCs can undergo tridermal differentiation, including hepatic specific lineages. Considering that iPSCs could be a source of hepatocyte regeneration, iPSC-based therapy has been widely implicated in the treatment of liver disease and hepatic regeneration. In the present review, we discuss the therapeutic potential of iPSCs in hepatic repair and focus on the clinical applications of iPSCs.
