Application of regorafenib and lapatinib in preparation of antitumor combination drug
A lapatinib and anti-tumor technology, which is applied in the field of anti-tumor drug preparation, can solve the problems such as the lack of combined use of regoratinib and lapatinib, and achieve the effects of superimposed curative effects, reduced dosage, and cost savings
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Embodiment 1
[0036] Example 1MTT method to detect the sensitivity of different tumor cells to rigatinib or lapatinib
[0037] Human colorectal cancer cells HCT116, SW620, HT29 and HCT8 were inoculated into 96-well plates with 5000-9000 cells per well. After the cells adhered to the wall, different concentrations of regoratinib or lapatinib were added respectively. of PBS solution. After culturing for 72 h, add 10 μL of MTT with a concentration of 5 mg / mL to each well and culture for another 4 h, then discard the culture solution and add 100 μL of DMSO to each well, and read the absorbance value of each well at 490 nm with a microplate reader. The half inhibitory concentration (IC50) was calculated by Bliss method. For specific results, see figure 1 .
[0038] From figure 1 It can be seen that the IC50 values of rigatinib and lapatinib on different tumor cells are in a relatively small range, and the IC50 values of rigatinib on different tumor cells are in the range of 15-25 μM , w...
Embodiment 2
[0039] Example 2 Effects of rigatinib or lapatinib alone and in combination on the survival rate of different tumor cells
[0040] Human colorectal cancer cells HCT116 and SW620 were inoculated into 96-well plates with 5000-9000 cells per well. After the cells adhered to the wall, PBS solutions with different ratios of rigatinib and lapatinib were added. . After culturing for 72 h, add 10 μL of MTT with a concentration of 5 mg / mL to each well and culture for another 4 h, then discard the culture solution and add 100 μL of DMSO to each well, and read the absorbance value of each well at 490 nm with a microplate reader. CompuSyn software was used for synergy comprehensive factor (Combinationindex, CI) analysis. see results figure 2 and Table 1, figure 2 It shows the cell viability of HCT116 and SW620 after combined treatment with different drug concentrations.
[0041]The CI value represents a composite index when the drug is combined, and the CI value is less than, equal ...
Embodiment 3
[0044] Example 3 Effects of rigatinib and lapatinib on the cell cycle of HCT116 and SW620 cells after separate and combined use
[0045] The cell cycle arrest of colorectal cancer cells HCT116 and SW620 induced by rigatinib and lapatinib alone and in combination was detected by flow cytometry. HCT116 and SW620 cells were divided into 3 × 10 per well 5 Cells were inoculated into 6-well plates at a density of 1000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000000 plate, after the cells adhered to the wall, the HCT116 and SW620 cells were divided into four experimental groups, and the following solutions were added respectively: the first group was the control group without treatment; Gritinib alone treatment group (3 μM, 10 μM, 30 μM three gradients); the third group was lapatinib single treatment group (3 μM, 10 μM, 30 μM three gradients); the fourth group was regatitinib Ni and lapatinib combined treatment group (co...
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