Application of natural compound Homoharringtonine in preparation of tumor treatment medicine
A tumor and tumor cell technology, applied in the direction of anti-tumor drugs, active ingredients of heterocyclic compounds, drug combinations, etc., can solve the problems of treatment failure and limited effect, and achieve the effect of promoting survival and good curative effect.
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Embodiment 1
[0046] Example 1, HHT inhibits the proliferation of non-small cell lung cancer and induces tumor cell apoptosis
[0047] 1. HHT inhibits the proliferation of non-small cell lung cancer
[0048] 1.1 HHT inhibits the proliferation of A549 cells
[0049] The experiment was repeated three times, and the specific steps of each repeated experiment were as follows:
[0050] A549 cells were divided into seven groups, each group 3 × 10 3 cells, these seven groups of A549 cells were processed according to the following method to obtain seven A549 cell culture fluids processed through different concentrations of HHT:
[0051] One group was in DMEM medium containing 10% fetal bovine serum in 5% CO 2 After culturing at 37°C for 20 hours in an incubator, add 10 μL of MTT aqueous solution with a concentration of 5 mg / mL and continue to cultivate for 4 hours, then suck out the medium containing MTT, add 150 μL DMSO, and place on a decolorization shaker at room temperature in the dark Shak...
Embodiment 2
[0098] Example 2, HHT induces apoptosis of non-small cell lung cancer cells through the mitochondrial apoptosis pathway
[0099] The experiment was repeated three times, and the specific steps of each repeated experiment were as follows:
[0100] Inoculate A549 cells into a 6-well plate containing DMEM medium, 1mL DMEM medium per well, 2×10 per well 5 A549 cells in 5% CO 2 Cultivate overnight at 37°C in an incubator to obtain the A549 cell culture solution after the cells adhere to the wall; add HHT to the A549 cells to obtain the HHT-A549 cell culture solution, and the concentration of HHT in the HHT-A549 cell culture solution is 4 μM; HHT-A549 cell culture medium in 5% CO 2 Incubate in an incubator at 37°C for 24 hours, discard the medium, wash the cells twice with 1×PBS, and then lyse the cells with RIPA lysis solution (RIPA lysis solution is a product of Beyond Biotechnology Co., Ltd.; product number P0013B) (each 200 μL RIPA lysate in the well), then centrifuged at 1,2...
Embodiment 3
[0106] Example 3, HHT inhibits STAT3 phosphorylation, thereby inhibiting STAT3 activation, from inhibiting cell survival
[0107] STAT3 expression levels and For its phosphorylation state, the primary antibody to detect the expression level of STAT3 is Anti-STAT3antibody[EPR787Y] (Abcam product, product number is ab68153), and the primary antibody to detect the phosphorylation state of STAT3 is Anti-STAT3(phosphoS727)antibody[E121-31 ] (Abcam product, product number is ab32143) and Anti-STAT3 (phosphoY705) antibody [EP2147Y] (Abcam product, product number is ab76315), the internal reference is GAPDH, and the primary antibody of GAPDH is Anti-GAPDHrabbitpolyclonalantibody (Sangon Bioengineering (Shanghai) Co., Ltd. Co., Ltd. products, the article number is D110016-0200), the results are as follows image 3 Shown in A.
[0108]The results showed that HHT inhibited the phosphorylation of STAT3Thr705, but did not affect the phosphorylation of STAT3Ser727 and the expression level...
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