agar and thiazolyl-blue

Titanium dioxide nanoparticles exhibit good anticancer and antibacterial activities. They are known to be environmentally friendly, stable, less toxic, and have excellent biocompatibility nature. Due to these properties, they are well suited for biological applications particularly in biomedical applications such as drug delivery and cancer therapy. In this research article, three medicinal herbs namely, Plectranthus amboinicus (Karpooravalli), Phyllanthus niruri (Keezhanelli), and Euphorbia hirta (Amman Pacharisi), were used to modify the surface of the TiO

Topics: Agar; Animals; Anti-Bacterial Agents; Antineoplastic Agents; Cell Line; Cell Survival; Euphorbia; HEK293 Cells; Humans; Hydrogen-Ion Concentration; Metal Nanoparticles; Mice; Microbial Sensitivity Tests; Models, Chemical; Nanotechnology; Phyllanthus; Plant Extracts; Plant Leaves; Plant Preparations; Plectranthus; Powders; Spectrophotometry, Ultraviolet; Spectroscopy, Fourier Transform Infrared; Tetrazolium Salts; Thiazoles; Titanium; X-Ray Diffraction

Oral squamous cell carcinoma (OSCC) is the most common type of oral cancer. Despite progress in the treatment of OSCC, overall survival has not improved substantially in the last three decades. Therefore, identification of reliable biomarkers becomes essential to develop effective anti-cancer therapy. In this study, we focused on the enzyme Nicotinamide N-methyltransferase (NNMT), which plays a fundamental role in the biotransformation of many xenobiotics. Although several tumors have been associated with abnormal NNMT expression, its role in cancer cell metabolism remains largely unknown. In this report, 7 human oral cancer cell lines were examined for NNMT expression by Real-Time PCR, Western blot and HPLC-based catalytic assay. Subsequently, we evaluated the in vitro effect of shRNA-mediated silencing of NNMT on cell proliferation. In vivo tumorigenicity of oral cancer cells with stable knockdown of NNMT was assayed by using xenograft models. High expression levels of NNMT were found in PE/CA PJ-15 cells, in keeping with the results of Western blot and catalytic activity assay. PE/CA PJ-15 cell line was stably transfected with shRNA plasmids against NNMT and analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and soft agar Assays. Transfected and control cells were injected into athymic mice in order to evaluate the effect of NNMT silencing on tumor growth. NNMT downregulation resulted in decreased cell proliferation and colony formation ability on soft agar. In athymic mice, NNMT silencing induced a marked reduction in tumour volume. Our results show that the downregulation of NNMT expression in human oral carcinoma cells significantly inhibits cell growth in vitro and tumorigenicity in vivo. All these experimental data seem to suggest that NNMT plays a critical role in the proliferation and tumorigenic capacity of oral cancer cells, and its inhibition could represent a potential molecular approach to the treatment of oral carcinoma.

Topics: Agar; Animals; Biomarkers, Tumor; Blotting, Western; Carcinoma; Cell Line, Tumor; Cell Proliferation; Chromatography, High Pressure Liquid; Female; Gene Silencing; Humans; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Mouth Neoplasms; Neoplasm Transplantation; Nicotinamide N-Methyltransferase; Real-Time Polymerase Chain Reaction; RNA Interference; RNA, Small Interfering; Tetrazolium Salts; Thiazoles

Increased levels of NF-κB are hallmarks of pancreatic ductal adenocarcinoma (PDAC) and both classical and alternative NF-κB activation pathways have been implicated.. Here we show that activation of the alternative pathway is a source for the high basal NF-κB activity in PDAC cell lines. Increased activity of the p52/RelB NF-κB complex is mediated through stabilization and activation of NF-κB-inducing kinase (NIK). We identify proteasomal downregulation of TNF receptor-associated factor 2 (TRAF2) as a mechanism by which levels of active NIK are increased in PDAC cell lines. Such upregulation of NIK expression and activity levels relays to increased proliferation and anchorage-independent growth, but not migration or survival of PDAC cells.. Rapid growth is one characteristic of pancreatic cancer. Our data indicates that the TRAF2/NIK/NF-κB2 pathway regulates PDAC cell tumorigenicity and could be a valuable target for therapy of this cancer.

Topics: Agar; Carcinoma, Pancreatic Ductal; Cell Line, Tumor; Cell Movement; Cell Nucleus; Cell Proliferation; Chemotaxis; Down-Regulation; Gene Expression Regulation, Neoplastic; Humans; NF-kappa B p52 Subunit; NF-kappaB-Inducing Kinase; Pancreatic Neoplasms; Proteasome Endopeptidase Complex; Protein Binding; Protein Serine-Threonine Kinases; Tetrazolium Salts; Thiazoles; TNF Receptor-Associated Factor 2

Ribosomal proteins are the components of ribosome, which also exhibit various secondary functions in DNA repair, apoptosis, drug resistance and proliferation. In our previous study of microarray, ribosomal protein L15 (RPL15) was identified as an upregulated gene in gastric cancer.. We investigated the expression of ribosomal protein L15 in gastric cancer and the effect of RPL15 on proliferation of gastric cancer.. It was found that the expression of RPL15 was markedly up-regulated in gastric cancer tissues. RPL15 was also highly expressed in gastric cancer cell lines AGS, MKN45, MKN28, SGC7901 and KATOIII. Inhibition of RPL15 expression by siRNA vector transfection suppressed the growth of SGC7901 cells significantly, which was independent of the expression of Cyclin D1 and B1. Down-regulation of RPL15 expression inhibited SGC7901 cell growth in soft agar and its tumorigenicity in nude mice.. RPL15 promotes cell proliferation and may be a potential target for anticancer therapy of gastric cancer.

Topics: Agar; Animals; Blotting, Western; Cell Line, Tumor; Cell Proliferation; Cyclin B; Cyclin B1; Cyclin D1; Down-Regulation; Flow Cytometry; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Oligonucleotide Array Sequence Analysis; Plasmids; Ribosomal Proteins; RNA, Small Interfering; Stomach Neoplasms; Tetrazolium Salts; Thiazoles; Transfection; Up-Regulation

hTid-1, a human homolog of the Drosophila tumor suppressor l(2)Tid and a novel DnaJ protein, regulates the activity of nuclear factor kappaB (NF-kappaB), but its mechanism is not established. We report here that hTid-1 strongly associated with the cytoplasmic protein complex of NF-kappaB-IkappaB through direct interaction with IkappaBalpha/beta and the IKKalpha/beta subunits of the IkappaB kinase complex. These interactions resulted in suppression of the IKK activity in a J-domain-dependent fashion and led to the cytoplasmic retention and enhanced stability of IkappaB. Overexpression of hTid-1 by using recombinant baculovirus or adenovirus led to inhibition of cell proliferation and induction of apoptosis of human osteosarcoma cells regardless of the p53 expression status. Adherent cultured cells transduced with Ad.hTid-1 detached from the dish surface. Morphological changes consistent with apoptosis and cell death were evident 48 h after Ad.EGFP-hTid-1 transduction. In contrast, cells transduced with Ad.EGFP or Ad.EGFP-hTd-1DeltaN100, a mutant that has the N-terminal J domain deletion and that lost suppressive activity on IKK, continued to proliferate. Similar data were obtained with A375 human melanoma cells. Ad.EGFP or Ad.EGFP-hTd-1DeltaN100 ex vivo-transduced A375 cells injected subcutaneously into nude mice produced growing tumors, whereas Ad.EGFP-hTid-1-transduced cells did not. Collectively, the data suggest that hTid-1 represses the activity of NF-kappaB through physical and functional interactions with the IKK complex and IkappaB and, in doing so, it modulates cell growth and death.

Topics: Adenoviridae; Agar; Animals; Apoptosis; Baculoviridae; Cell Death; Cell Line; Cell Line, Tumor; Cell Proliferation; Cytoplasm; Drosophila; Gene Deletion; Gene Expression Regulation; Genes, Reporter; Glutathione Transferase; Green Fluorescent Proteins; Heat-Shock Proteins; HSP40 Heat-Shock Proteins; Humans; I-kappa B Proteins; Immunoprecipitation; Lentivirus; Male; Mice; Mice, Nude; Microscopy, Fluorescence; Mutation; Neoplasm Transplantation; NF-kappa B; Osteosarcoma; Plasmids; Protein Binding; Protein Structure, Tertiary; Recombinant Fusion Proteins; Signal Transduction; Tetrazolium Salts; Thiazoles; Time Factors; Transfection; Tumor Suppressor Protein p53

RWJ-241947 (MCC-555) is a novel peroxisome proliferator-activated receptor-gamma ligand of the thiazolidinedione class that was recently developed as an antidiabetic drug with unique properties. Some thiazolidinediones have anticancer activity against solid and hematological malignancies; the anticancer potency of RWJ-241947 has not been examined. We, therefore, investigated these effects in vitro and in vivo either alone or in combination with other compounds.. Tumor growth was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, soft agar colony assay in vitro, and xenografts in nude mice. Its effects on cell cycle, differentiation, and apoptosis were examined.. In vitro studies using various solid and hematological tumor cell lines showed that RWJ-241947 had antiproliferative activity against prostate cancer cells, with the strongest effect against the androgen-independent PC-3 prostate cancer cells. It increased expression of cyclin-dependent kinase inhibitor p21(WAF1), deceased cyclin E, and induced apoptosis in PC-3 cells. It increased E-cadherin and lowered protein expression of prostate-specific antigen without down-regulating the androgen receptor in androgen-dependent LNCaP prostate cancer cells. Reporter gene assays showed that this peroxisome proliferator-activated receptor-gamma ligand inhibited androgen activation of the androgen receptor response elements of the prostate-specific antigen gene. Remarkably, in vivo treatment of male beige/nude/X-linked immunodeficient (BNX) mice with RWJ-241947 profoundly suppressed growth of PC-3 prostate cancer xenografts with prominent apoptosis, as well as fibrosis, including inflammatory and giant cell reaction in the remaining tumor tissue. Notably, the experimented mice had a significantly decreased cholesterol. In addition, we studied the combination of arsenic trioxide (As2O3), which is used in the treatment of multiple myeloma, and RWJ-241947; these two reagents together prominently inhibited proliferation and caused apoptosis of multiple myeloma cells.. RWJ-241947 has surprisingly potent antiproliferative effects against prostate cancer cells in vivo, and it enhances the antitumor activity of As2O3 against myeloma cells. Small, well-defined clinical studies using RWJ-241947 are in order for these cancers.

Topics: Agar; Animals; Antineoplastic Agents; Apoptosis; Arsenic Trioxide; Arsenicals; Blotting, Western; Caspases; CD36 Antigens; Cell Cycle; Cell Differentiation; Cell Division; Cell Line, Tumor; Cell Survival; Genetic Linkage; Humans; Immunologic Deficiency Syndromes; Ligands; Luciferases; Male; Mice; Multiple Myeloma; Oxides; Prostatic Neoplasms; Receptors, Cytoplasmic and Nuclear; Tetrazolium Salts; Thiazoles; Thiazolidinediones; Time Factors; Transcription Factors; Transfection; U937 Cells; X Chromosome

The small GTPase RhoA has been implicated in the regulation of cell morphology, motility, and transformation, but the role of RhoA protein in the carcinogenesis of gastric cancer remains unclear. In the present study, we have analyzed the expression status of the RhoA protein in human gastric cancer cells and tissues and investigated the possible involvement of RhoA in regulating the malignant phenotype of gastric cancer cells.. RhoA expression was analyzed by immunohistochemistry and Western blot in gastric cancer tissues and cell lines. The RhoA-specific small interfering RNA (siRNA) vector was designed and constructed. We examined the role of RhoA in the malignant phenotype of gastric cancer cells by using siRNA knockdown and dominant-negative RhoA mutant suppression of endogenous RhoA activity.. RhoA was found frequently overexpressed in gastric cancer tissues and cells compared with normal tissues or gastric epithelial cells. RhoA-specific siRNA could specifically and stably reduce RhoA expression up to 90% in AGS cells. Both RhoA-specific siRNA and dominant-negative RhoA expressions could significantly inhibit the proliferation and tumorigenicity of AGS cells and enhance chemosensitivity of the cancer cells to Adriamycin and 5-fluorouracil.. RhoA may play a critical role in the carcinogenesis of gastric cancer, and the interference of RhoA expression and/or activity could provide a novel avenue in reversing the malignant phenotype of gastric cancer cells.

Topics: Agar; Animals; Antibiotics, Antineoplastic; Antimetabolites, Antineoplastic; Antineoplastic Agents; Blotting, Western; Cell Cycle; Cell Differentiation; Cell Line, Transformed; Cell Line, Tumor; Cell Movement; Cell Proliferation; Disease Progression; DNA; Dose-Response Relationship, Drug; Doxorubicin; Epithelial Cells; Flow Cytometry; Fluorouracil; Genes, Dominant; Humans; Immunohistochemistry; Mice; Mutation; Neoplasm Invasiveness; Neoplasm Metastasis; Oligonucleotides; Phenotype; rhoA GTP-Binding Protein; RNA, Small Interfering; Stomach Neoplasms; Tetrazolium Salts; Thiazoles; Time Factors; Transfection; Wound Healing

Topics: Agar; Animals; Antineoplastic Agents; Cell Count; Cell Death; Cell Division; Cell Line; Cell Survival; Clone Cells; DNA; DNA Damage; DNA Topoisomerases, Type II; Tetrazolium Salts; Thiazoles; Topoisomerase II Inhibitors