Application of grape seed procyanidine in preparation of combination medicine in cancer chemotherapy scheme
A technology of proanthocyanidins and chemotherapeutic drugs, which is applied in the medical field, can solve the problems of cell drug resistance and cannot fundamentally reduce cell drug resistance, and achieve the effect of avoiding the increase of drug dose, reducing risk and reducing dose
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Embodiment 1
[0028] Example 1: Analysis of protein expression in HL-60 cells and HL-60 / ADR cells
[0029] This example is an experiment for analyzing protein expression in HL-60 cells and HL-60 / ADR cells, and the experimental method is as follows.
[0030] 1.1 Extraction of total cell protein
[0031] 1.1.1 Instrument: high-speed / refrigerated centrifuge (Germany eppendorf, 5415D)
[0032] 1.1.2 Reagents:
[0033] (1) Whole protein lysate (Shanghai Kangcheng Bioengineering Co., Ltd.)
[0034] (2) Protease inhibitor mixture (Shanghai Kangcheng Bioengineering Co., Ltd.)
[0035] (3) PMSF solution (Shanghai Kangcheng Biological Engineering Co., Ltd.)
[0036] (4) Phosphatase Mixture (Shanghai Kangcheng Biological Engineering Co., Ltd.)
[0037] 1.1.3 Solution preparation
[0038] Protein extraction reagents:
[0039]
[0040] 1.1.4 Cell collection (HL-60 and HL-60 / ADR cells are suspension cells)
[0041] (1) HL-60 and HL-60 / ADR cells in 2×10 6 Inoculate in a 6cm culture dish. After...
Embodiment 2
[0081] Example 2: mRNA level analysis in HL-60 cells and HL-60 / ADR cells
[0082] This example is an experiment for analyzing protein expression in HL-60 cells and HL-60 / ADR cells, and the experimental method is as follows.
[0083] 2.1 Trizol extraction of total cellular RNA (050403A)
[0084] 2.1.1 Instruments and materials: high-speed / refrigerated centrifuge (Germany eppendorf, 5415D)
[0085] 2.1.2 Reagents:
[0086] (1) Trizol (2) DEPC water (3) Chloroform (4) Isopropanol (5) Ethanol
[0087] 2.1.3 Experimental steps
[0088] (1) Cell collection: a. HL-60 and HL-60 / ADR cells in 2×10 6 Inoculate in a 6cm culture dish. After the cells have grown for 48 hours, carefully transfer the cells into a 15ml centrifuge tube, centrifuge at 1500 rpm for 10 minutes to sediment the suspended cells, and discard the supernatant;
[0089] (2) Add 1ml PBS to the centrifuge tube, transfer the resuspended cells to a 1.5ml tube, centrifuge at 1500 rpm for 5 minutes and discard the superna...
Embodiment 3
[0147] Example 3: Analysis of the IC50 of 8 different drugs in HL-60 cells and HL-60 / ADR cells and the effect of grape seed proanthocyanidins on drug resistance reversal
[0148] This example is an analysis experiment of the IC50 of 8 different drugs in HL-60 cells and HL-60 / ADR cells and the effect of grape seed proanthocyanidins (hereinafter referred to as GSPE) in reversing drug resistance.
[0149] The 8 middle medicines shown in the present embodiment are used clinically commonly used cancer chemotherapeutic drugs, are respectively: doxorubicin (hereinafter denoted as ADR), daunorubicin (hereinafter denoted as DNR), etoposide (hereinafter denoted as VP16), vincristine (hereinafter referred to as VCR), cytarabine (hereinafter referred to as Ara-C), homoharringtonine (hereinafter referred to as HHT), mitoxantrone (hereinafter referred to as MTZ), pyridine Rubicin (hereinafter referred to as THP). The experimental method is as follows.
[0150] 3.1 Cell Treatment
[0151]...
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