Neoplastic growth could arise from different cell types within the body, creating a spectrum of disease types that vary in pathological behaviour and treatment approaches.
The development of cancer arises from a dysregulation in cellular proliferation, which usually arises from inappropriate responses to external stimuli. Through the years, discovery of mutations in key oncogenes and tumour suppressor genes has been pivotal in characterising biological aberrancies that favour oncogenesis. For instance, many cancers harbour activating mutations in components of several survival pathways (e.g. PI3K and MAPK pathways) leading to constitutive signalling. Downstream effects include increased production of growth factors for inter and intracellular interactions, signalling cross talks and loss of response to inhibitory stimuli. The acquisition of cancer hallmarks, as outlined by Hanahan and Weinberg (2011), has been the foundation of studying cancer biology and determining various approaches for treatment purposes. For years, therapeutic regimes have been developed to target the following oncogenic drivers:
- Sustaining proliferative signalling
- Resisting cell death
- Inducing angiogenesis
- Enabling replicative immortality
- Activating invasion and metastasis
- Evading growth suppressors
Cellomatics provides expertise in generating preclinical data for clients within the oncology area of interest. Alongside the ongoing development of disease models, our team routinely performs high throughput drug screens followed by a wide range of assays to measure certain biological responses that complement the hallmarks of cancer.
Neutrophil migration through the HUVEC monolayer
A statistically significant increase in neutrophil migration towards the fMLP chemoattractant was observed in HUVEC monolayers treated with TNFα when compared to untreated HUVEC monolayers (ns=not significant; **p<0.01; ***p<0.001±SEM).
HLF (human lung fibroblasts) cells were seeded at a density of 4000 cells/well and incubated overnight at 37oC in a humidified incubator. The cells were treated with PDGF-BB (3ng/mL; 10ng/mL) and Mitomycin C (10µM; 100µM). The BrdU assay showed that treatment with PDGF-BB and 10% FBS resulted in a significant increase in the cell proliferation, as indicated by the increase in the absorbance values. Treatment with Mitomycin C significantly reduced the absorbance, suggesting an inhibition in cell proliferation (***p<0.001 ± SEM).
HMC1.2 cells were treated with a (or the) test compound at 6 different concentrations. The % cell viability post-compound-treatment was determined by means of an MTT assay. A significant reduction in cell viability was observed with δ-Tocopherol when compared to the positive and vehicle controls (***p<0.001; n=5; ±SEM)
Caspase 3/7 Assay
HeLa cells were treated with Staurosporine for 4 hours. The Caspase-Glo 3/7 Reagent was added directly to cells in 96-well plates and incubated at room temperature for 30 minutes before recording luminescence (RLU). Each point represents the average of 3 replicates (error bars represent ±SEM).
A549 cells were treated with drug compounds at 8 different concentrations. Cell Viability and cytotoxic responses were assessed using the CellTitre-Glo and LDH assays. Staurosporine (positive control compound) induced a significant reduction in cell viability when compared to untreated or vehicle control (***p<0.001; ±SEM).
Dose Response for Talazoparib and Minocycline of PARP activity and Proliferation in UWB1.289
The ovarian cancer cell line UWB1.289 was treated for 24 hours with increasing concentrations of Talazoparib and Minocycline. PARP activity was measured by an in vitro cell free enzymatic assay. Proliferation was assessed by BrdU incorporation (n=3±SEM).
3D spheroid models
MDA-MB436 cells cultured in 3D cell culture media were allowed to form spheroids 5 days prior to Talazoparib and Bleomycin drug treatment. Following the drug treatment, spheroids were stained with DEAD Red and SYTO Green fluorescent dye every 72 hours. Spheroid growth and morphology were assessed using JuLITM Stage Automated cell imaging system to determine the effect of drug treatments. Combination of Talazoparib and Bleomycin resulted in an increase in cell death and an observable loss of spheroids.
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