ABSTRACT
BACKGROUND/AIM: Currently, treatment of non-muscle invasive bladder cancer causes significant deterioration in a patient's quality of life (QOL). Therefore, development of novel therapeutic options without the deterioration of QOL is very important. In this study, we assessed the anti-tumor effect of lentivirus-mediated gene transfection of tumor-suppressor genes in human bladder cancer cells. MATERIALS AND METHODS: Lentiviral vectors that contained the tumor suppressor genes, p53, p16, and PTEN, were transfected into human bladder cancer cell lines, 5637, T24, 253J, and UMUC3, and the normal human uroepithelial cell line, SV-HUC-1. RESULTS: Significant growth inhibition was observed in bladder cancer cells on transfection with the p16 and PTEN vectors. However, the effect of the p53 vector was limited. In normal cells, the lentiviral vectors did not exhibit a significant growth inhibitory effect. CONCLUSION: Lentiviral vector-mediated gene transfection is useful for the application of gene therapy in bladder cancers.
Subject(s)
Genes, Tumor Suppressor , Genetic Therapy/methods , Lentivirus/genetics , Transfection/methods , Urinary Bladder Neoplasms/genetics , Urinary Bladder Neoplasms/therapy , Cell Line, Tumor , Cyclin-Dependent Kinase Inhibitor p16/biosynthesis , Cyclin-Dependent Kinase Inhibitor p16/genetics , Genes, p16 , Genes, p53 , Genetic Vectors , Humans , Tumor Suppressor Protein p53/biosynthesis , Tumor Suppressor Protein p53/genetics , Urinary Bladder Neoplasms/virologyABSTRACT
AIM: Naftopidil is used to treat benign prostate hyperplasia. Moreover, previous studies have shown that naftopidil reduced viability of many types of cancer cells. Therefore, we investigated the antitumor mechanism of naftopidil in this study. MATERIALS AND METHODS: We used the HGC27 human gastric cancer cell line. It was treated with naftopidil, pan-caspase inhibitor, and chloroquine diphosphate (CQ). Cell viability and cell death were investigated by the assay and annexin V/ propidium iodide assay. Phosphorylation of protein kinase B (AKT) (Ser473) was measured by western blotting. Alteration of light chain 3B (LC3B) was investigated by western blotting and immunofluorescence. RESULTS: Naftopidil reduced phospho-AKT (Ser473) and altered LC3B. Combination of naftopidil and CQ reduced cell viability and phospho-AKT (Ser 473). CONCLUSION: Naftopidil induces apoptosis and autophagy of HGC27 cells, however, autophagy is considered to inhibit apoptosis. We concluded naftopidil and CQ have a synergistic antitumor effect.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/pharmacology , Apoptosis/drug effects , Autophagy/drug effects , Naphthalenes/pharmacology , Piperazines/pharmacology , Stomach Neoplasms/drug therapy , Cell Line, Tumor , Chloroquine/administration & dosage , Chloroquine/analogs & derivatives , Chloroquine/pharmacology , Drug Synergism , Humans , Microtubule-Associated Proteins/metabolism , Naphthalenes/administration & dosage , Phosphorylation , Piperazines/administration & dosage , Proto-Oncogene Proteins c-akt/metabolism , Stomach Neoplasms/metabolism , Stomach Neoplasms/pathologyABSTRACT
A new cell line of human uterine endometrial undifferentiated carcinoma, designated as TMG-L, was established from the metastatic lymph node of 56-year-old patient TMG-L cells have been cultured with Ham's F-12 medium supplemented with 10% FCS and grew as a loosely adherent monolayer with polygonal or spindle-shaped cells exhibiting poor cell-cell contact and piled up against each other, showing a tendency to grow as floating cells. The doubling time of this cell line was about 48 hours, and chromosomal analysis revealed aneuploidy at passage 25. The cells formed tumors in SCID mouse, the histology of which was similar to that of undifferentiated carcinoma component of primary tumor. TMG-L cells showed the loss of expression and membranous localization of either E-cadherin or alpha-catenin, implied corresponding loss of their adhesive function. And this dysfunction implicated the biological aggressive behavior of uterine endometrial undifferentiated carcinoma. This cell line appears to provide a useful system for studying uterine undifferentiated carcinoma in vivo and in vitro.