ABSTRACT
Dengue fever is a mosquito-borne disease that has rapidly spread throughout the last few decades. Most preventive mechanisms to deal with the disease focus on the eradication of the vector mosquito and vaccination campaigns. However, appropriate mechanisms of response are indispensable to face the consequent events when an outbreak takes place. This study applied single and multiple objective linear programming models to optimize the allocation of patients and additional resources during an epidemic dengue fever outbreak, minimizing the summation of the distance travelled by all patients. An empirical study was set in Ciudad del Este, Paraguay. Data provided by a privately run health insurance cooperative was used to verify the applicability of the models in this study. The results can be used by analysts and decision makers to solve patient allocation problems for providing essential medical care during an epidemic dengue fever outbreak.
ABSTRACT
BACKGROUND: In vitro modelling of cancer cells is becoming more complex due to prevailing evidence of intimate interactions between cancer cells and their surrounding stroma. A co-culture system which consists of more than one cell type is physiologically more relevant and thus, could serve as a useful model for various biological studies. An assay that specifically detects the phenotypic changes of cancer cells in a multi-cellular system is lacking for nasopharyngeal carcinoma (NPC). RESULTS: Here, we describe a luciferase/luciferin (XenoLuc) assay that could specifically measure changes in the proliferation of cancer cells in the co-culture system using two modified NPC patient-derived tumour xenograft (PDTXs) cells: Xeno284-gfp-luc2 and XenoB110-gfp-luc2. Through this assay, we are able to show that the growth of NPC xenograft cells in both two-dimensional (2D) and three-dimensional (3D) models was enhanced when co-cultured with normal human dermal fibroblasts (NHDFs). In addition, potential applications of this assay in in vitro drug or inhibitor screening experiments are also illustrated. CONCLUSIONS: XenoLuc assay is specific, sensitive, rapid and cost-effective for measuring the growth of luciferase-expressing cells in a co- or multiple-culture system. This assay may also be adapted for tumour microenvironment studies as well as drug screening experiments in more complex 3D co-culture systems.
Subject(s)
Fibroblasts/metabolism , Green Fluorescent Proteins/metabolism , Luciferases/metabolism , Nasopharyngeal Carcinoma/metabolism , Animals , Cell Line, Tumor , Cells, Cultured , Coculture Techniques , Fibroblasts/cytology , Green Fluorescent Proteins/genetics , HEK293 Cells , Humans , Luciferases/genetics , Luminescent Measurements/methods , Mice, Inbred NOD , Mice, Knockout , Mice, SCID , Nasopharyngeal Carcinoma/genetics , Nasopharyngeal Carcinoma/pathology , Transplantation, HeterologousABSTRACT
Copper(II) ternary complex, [Cu(phen)(C-dmg)(H2O)]NO3 was evaluated against a panel of cell lines, tested for in vivo efficacy in nasopharyngeal carcinoma xenograft models as well as for toxicity in NOD scid gamma mice. The Cu(II) complex displayed broad spectrum cytotoxicity against multiple cancer types, including lung, colon, central nervous system, melanoma, ovarian, and prostate cancer cell lines in the NCI-60 panel. The Cu(II) complex did not cause significant induction of cytochrome P450 (CYP) 3A and 1A enzymes but moderately inhibited CYP isoforms 1A2, 2C9, 2C19, 2D6, 2B6, 2C8 and 3A4. The complex significantly inhibited tumor growth in nasopharyngeal carcinoma xenograft bearing mice models at doses which were well tolerated without causing significant or permanent toxic side effects. However, higher doses which resulted in better inhibition of tumor growth also resulted in toxicity.
Subject(s)
Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Carcinoma/drug therapy , Copper/chemistry , Nasopharyngeal Neoplasms/drug therapy , Organometallic Compounds/chemistry , Organometallic Compounds/pharmacology , Xenograft Model Antitumor Assays , Animals , Antineoplastic Agents/toxicity , Carcinoma/pathology , Cell Line, Tumor , Cytochrome P-450 Enzyme Inhibitors/chemistry , Cytochrome P-450 Enzyme Inhibitors/pharmacology , Cytochrome P-450 Enzyme Inhibitors/toxicity , Cytochrome P-450 Enzyme System/biosynthesis , Cytochrome P-450 Enzyme System/metabolism , Dose-Response Relationship, Drug , Enzyme Induction/drug effects , Female , Hepatocytes/drug effects , Mice , Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms/pathology , Organometallic Compounds/toxicity , RatsABSTRACT
Subpopulations of nasopharyngeal carcinoma (NPC) contain cells with differential tumourigenic properties. Our study evaluates the tumourigenic potential of CD24, CD44, EpCAM and combination of EpCAM/CD44 cells in NPC. CD44br and EpCAMbr cells enriched for higher S-phase cell content, faster-growing tumourigenic cells leading to tumours with larger volume and higher mitotic figures. Although CD44br and EpCAMbr cells significantly enriched for tumour-initiating cells (TICs), all cells could retain self-renewal property for at least four generations. Compared to CD44 marker alone, EpCAM/CD44dbr marker did not enhance for cells with faster-growing ability or higher TIC frequency. Cells expressing high CD44 or EpCAM had lower KLF4 and p21 in NPC subpopulations. KLF4-overexpressed EpCAMbr cells had slower growth while Kenpaullone inhibition of KLF4 transcription increased in vitro cell proliferation. Compared to non-NPC, NPC specimens had increased expression of EPCAM, of which tumours from advanced stage of NPC had higher expression. Together, our study provides evidence that EpCAM is a potentially important marker in NPC.
Subject(s)
CD24 Antigen/metabolism , Epithelial Cell Adhesion Molecule/metabolism , Hyaluronan Receptors/metabolism , Nasopharyngeal Carcinoma/metabolism , Nasopharyngeal Neoplasms/metabolism , Neoplastic Stem Cells/metabolism , Transplantation, Heterologous , Animals , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , CD24 Antigen/genetics , Cell Line, Tumor , Epithelial Cell Adhesion Molecule/genetics , Female , Humans , Hyaluronan Receptors/genetics , Kruppel-Like Factor 4 , Mice, Inbred NOD , Mice, Knockout , Mice, SCID , Nasopharyngeal Carcinoma/genetics , Nasopharyngeal Carcinoma/pathology , Nasopharyngeal Neoplasms/genetics , Nasopharyngeal Neoplasms/pathologyABSTRACT
The molecular events that drive the progression of Epstein-Barr virus (EBV)-associated nasopharyngeal carcinoma (NPC) are still to be elucidated. Here, we report for the first time the pathogenic significance of an NPC-associated gene, wingless-type MMTV integration site family, member 5A (WNT5A) and the contribution of EBV to its expression. WNT5A is a representative Wnt protein that activates non-canonical Wnt signalling. With regard to its role in carcinogenesis, there is conflicting evidence as to whether WNT5A has a tumour-promoting or tumour-suppressive role. We show that WNT5A is upregulated in primary NPC tissue samples. We also demonstrate that WNT5A expression was dramatically increased in NPC cell lines expressing the EBV-encoded LMP2A gene, suggesting that this EBV-encoded latent gene is responsible for upregulating WNT5A in NPC. In addition, in vitro WNT5A overexpression promotes the proliferation, migration and invasion of NPC cells. Our results not only reveal pro-tumorigenic effects of WNT5A in NPC but also suggest that WNT5A could be an important therapeutic target in patients with EBV-associated disease.
