Application of scarlet 808 in preparation of tumor multidrug resistance reversal agent
A multi-drug resistance, multi-drug resistance technology, applied in the treatment of multi-drug resistant tumors, the field of medicine
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0021] Embodiment 1: MTT experiment
[0022] MTT method is also called MTT colorimetric method. MTT assay was used to detect the proliferation inhibitory activity of Scarlet 808 on parental cells (parent plant) and drug-resistant cells (Resistant cells). Take the logarithmic growth parental cells and the corresponding drug-resistant cells, discard the old medium, and gently wash the cells with preheated PBS, discard the PBS, add trypsin and digest at 37°C for 1 min. After the digestion is complete, add pre-warmed complete DMEM medium to stop the digestion and collect into a 50 mL centrifuge tube, then count the cells. Cells were diluted to 2.5×10 4 Cells / mL, spread evenly into 96-well plates at a volume of 200 μL per well (the number of cells per well is 5000), and culture in an incubator for 6 h to make them adhere to the wall. After the cells adhered to the wall, a certain concentration gradient of Scarlet 808 was added, and in the presence of 5% CO 2 After culturing in ...
Embodiment 2
[0023] Example 2: Reversal experiment
[0024] Take parental cells and drug-resistant cells that are in logarithmic growth phase and in good condition, and seed the plate with 5000 cells per well, and the medium content in each well is 200 μL. After culturing for 6 h to adhere to the wall, suck away 40 μL of medium, and add a certain concentration of Scarlet 808. After continuing to culture for two hours, add 20 μL of chemotherapeutic drugs diluted according to the appropriate concentration gradient, add MTT after 72 h of culture, and detect according to the above method. Drug resistance multiple calculation method: IC 50 Drug-resistant cells / IC 50 Parental cell control group. The results are shown in Table 1-5, Scarlet 808 can reduce the IC of drug-resistant cells 50 , and had no effect on parental cells.
[0025] Table 1 Effect of scarlet 808 combined with chemotherapy drugs on the proliferation of KB-3-1 and KB-C2 cells
[0026] a RF: Resistant Fold, resistance multi...
Embodiment 3
[0040] Example 3: Using Flow Cytometry to Detect the Effect of Scarlet 808 on the Accumulation Level of Chemotherapeutic Drugs in Tumor Multidrug Resistant Cells Using PhA and ADR as Fluorescently Labeled Substrates to Study the Effect of Scarlet 808 on Two Drug Resistance Proteins in Tumor Multidrug Resistant Cells Effects of accumulation in resistant cells. Take the cells in the logarithmic growth phase and in good condition, trypsinize, centrifuge and resuspend, and count the cells to prepare a final concentration of 1×10 4 cells / mL of cell suspension was evenly spread in 24-well plates at 1 mL per well (10,000 cells per well), and the next drug treatment was carried out after 6 h of attachment. The orifice plate was set up with parental cells KB-3-1, NCI-H460 and S1 as negative controls, and positive reversal agent controls as ABCB1-specific reversal agent verapamil and ABCG2-specific reversal agent Ko143, respectively. First, add scarlet 808 (final concentration of 1 μM ...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More