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Mitochondrial activity inhibitors of cancer-initiating cells and use thereof
An active inhibitor and starting cell technology, which can be used in the fields of organic active ingredients, detection of programmed cell death, and medical preparations containing active ingredients, which can solve the problem of no effective treatment for gliomas.
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Embodiment 1
[0221] Example 1: Demonstration of oxidative cellular energy production processes and cancer initiation FL1 + Analysis of Mitochondrial Activity in Cell Populations
[0222] figure 1 Anaerobic and aerobic pathways in cells are shown. Glycolysis is a process that metabolizes glucose to pyruvate in the cytoplasm. Under hypoxic conditions, pyruvate is converted to lactate by LDH: 1GLC→2ATP.
[0223] Tricarboxylic acid (TCA) combined oxidative phosphorylation (OXPHOS) is the process of generating ATP from ADP using pyruvate obtained from glycolysis, electron transfer via NADH and FADH2 to respiratory chain complexes in mitochondria, and proton gradient pumps. Under aerobic conditions: 1GLC → 36ATP.
[0224] Study metabolic pathways in cancer-initiating cells by measuring the following parameters:
[0225] Number of active mitochondria (eg M75-13): cells were harvested, dissociated, washed and treated with M75 diluted at a final concentration of 250 nM in DMEM-F12-Glutam...
Embodiment 2
[0237] Example 2: Testing Potential Anti-Cancer Stem Cell Agents (Inhibitors)
[0238] According to the results of mitochondrial activity, metabolite content and inhibitor effect of anaerobic energy production pathway (described in Example 1), the FL1 under the aerobic glycolysis system 0 or FL1 - cells compared to FL1 + Cells likely preferentially use the aerobic pathway to generate their own energy. Therefore, any agent with a potent ability to inhibit mitochondrial activity would impair energy production in the CIC, which would likely kill the CIC. This was tested in Figure 5 and in the in vitro recurrence assay described below:
[0239] For short-term / dose response (48 hours): Dissociated gliomaspheres, attached glioma and normal cells were plated at 10 cells / μl on DMEM-F12 Glutamax, BIT20% or B27 (1 / 50 ), Penicinil / Streptomycin 1 / 1000, 1 ng / ml of reduced mitogen or reduced level of serum (2.5%) was used.
[0240] For long-term treatment / recovery assays (T10 and / or ...
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