Anticancer pharmaceutical composition containing IF1 as active ingredient
A technology of active ingredients and compositions, which is applied in the field of anticancer drug compositions containing IF1 as an active ingredient, and can solve problems such as no treatment method and severe toxicity
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Embodiment 1
[0066] Example 1: Measurement of extracellular ATP release
[0067] Breast cancer (MDA-MB-231, MCF7, T47D), liver cancer (HepG2) and cervical cancer (HeLa) cell lines were cultured in DMEM medium containing 10% FBS and 1% penicillin / streptomycin under 5% carbon dioxide Incubate in a 37 °C incubator.
[0068] Extracellular ATP release after treatment with IF1 was measured using the CellTiter-Glo Luminescence Assay Kit (Promega, Madison, WI, USA). HepG2 (liver cancer cell line) or MDA-MB-231 (breast cancer cell line) cells (3×10 4 cells / well) were treated with IF1 (100ng / mL), and then incubated at 37°C and 5% CO 2 Incubate in 1% BSA / DMEM medium in an incubator for 10 seconds, 30 seconds, 1 minute, 5 minutes, 10 minutes and 30 minutes under the conditions of . Each medium was then harvested and subjected to luminescence assays.
[0069] As a result, it can be seen that after treatment with IF1, the ATP concentration in the medium rapidly increased ( figure 1 ). It can be se...
Embodiment 2
[0070] Example 2: In Vitro Cancer Cell Proliferation and Death Assays
[0071] To investigate the cytotoxic and proliferation-inhibitory effects of IF1 in cancer cells, MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) assay and CVS (Crystal Violet Solution) assay. MTT and CVS assays are methods for measuring the growth of living cells. MTT uses MTT reduction by the mitochondrial reductase of living cells, while CVS measures the viability of cells using the phenomenon in which cells detach from the plate after death, whereby the level of crystal violet staining decreases.
[0072] Each cancer cell line (HepG2, MDA-MB-231, T47D, HeLa) was seeded into a 96-well plate and incubated at 37°C and 5% CO 2 Incubate overnight in the incubator under the same conditions, and then again overnight in fresh serum-free medium the next day. In addition to the control group, cells (MDA-MB-231 cell line) were treated with various concentrations of IF1 at 6h, 24h and 48h. Aft...
Embodiment 3
[0076] Example 3: Inhibitory effect of treatment with IF1 on cancer cell migration
[0077] Cancer cells in the human body migrate to healthy tissues and cause metastasis through their characteristics and the composition of the microenvironment. Metastasis of cancer cells means the transformation of normal tissues into cancerous tissues. Therefore, the effect of IF1 on cancer cells was observed by cell migration assay.
[0078] Each well was scraped in one direction, then MDA-MB-231 cells cultured in a 6-well plate were treated with different concentrations of IF1 and observed with a light microscope at 48 hours.
[0079] The results showed that as the concentration of IF1 increased, the number of cells migrating to the empty space decreased, suggesting that IF1 inhibited the migration of cancer cells ( image 3 ).
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