Pharmaceutical composition for treating cancer and application of pharmaceutical composition
A composition and drug technology, applied in the field of pharmaceutical compositions for the treatment of cancer, can solve problems such as liver damage and bone marrow suppression, and achieve the effect of reducing toxicity, reducing damage, making and administering conveniently and quickly
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Embodiment 1
[0063] Example 1, the inhibitory effect of methotrexate and JQ-1 inhibitors on the growth of human pancreatic cancer cell Panc1 cells
[0064] Cell proliferation detected by CCK-8 method: Panc1 cells were inoculated with 10% fetal bovine serum, 1×10 5 U / L penicillin and 100g / L streptomycin in DMEM medium, 5% CO 2 Incubator at 37°C. Panc1 cells in the logarithmic growth phase were divided into 3 × 10 3 and 5×10 3 seeded in 96-well plates. After culturing for 24 hours, 0, 10, 20, 40, 100, 200, 400 nM of MTX and 0, 125, 250 nM of JQ-1 inhibitor were added. Four replicate wells were set up in each group, and a negative control was set up at the same time. placed in 5% CO 2 Incubate at 37°C for 24h and 48h. Add 10 μL of CCK-8 solution to each well, continue to incubate in the cell culture incubator for 1 hour, and measure the absorbance at 450 nm. Calculate the inhibition rate according to the following formula:
[0065] Survival rate (%)=[(A450, Sample-A450, Blank) / (A450,...
Embodiment 2
[0070] Example 2, methotrexate and JQ-1 inhibitors induce cell death and autophagy in human pancreatic cancer cells Panc1
[0071] In this example, the concentration of MTX used is 100nM / L, and the relative molecular mass is 454.44g / mol; the concentration of JQ-1 inhibitor is 200nM / L, and the relative molecular mass is 456.99g / mol.
[0072] How to use MTX: (1) Dissolve MTX in DMSO to make 10mM stock solution; (2) Dilute to 100nmol / L with cell culture medium and add to cell culture dish.
[0073]
[0074] JQ-1 inhibitor usage method: (1) JQ-1 inhibitor is dissolved in DMSO, and prepared as a 10mM stock solution; (2) used for cell culture and diluted to 200nmol / L, and added to a cell culture dish.
[0075]
[0076] Combined use of MTX+JQ-1 inhibitor: Take appropriate amount of stock solution of JQ-1 and MTX and dilute to a specific concentration with culture medium (JQ-1 inhibitor 200nM, MTX 100nM).
[0077] AnnexinV-PE / 7-AAD double staining combined with flow cytometry t...
Embodiment 3
[0087] Example 3, MTX and JQ-1 inhibitors jointly affect the folic acid, purine and pyrimidine synthesis pathways of pancreatic cancer cells
[0088] The gene chip clarified the cellular processes affected by JQ-1 inhibitors: treated Panc1 cells with JQ-1 inhibitors and DMSO (control), extracted RNA for gene chip analysis; used bioinformatics methods to find differentially expressed genes and analyzed them Pathway analysis. See Figure 14 and Figure 15 , as can be seen from the figure, the bioinformatics method screened out the 100 most significantly changed differential genes after JQ-1 inhibitor treatment, GO and KEGG pathway analysis showed that the synthesis of purine and pyrimidine was significantly affected after JQ-1 inhibitor treatment .
[0089] Real-time fluorescent quantitative PCR detection of key gene changes in folic acid, purine and pyrimidine synthesis pathways: Panc1 cells were treated with MTX and JQ-1 inhibitors, RNA was extracted, and real-time fluoresc...
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