Application of fulvestrant in preparing antimicrotubular chemotherapeutic drug resistance reversal agent
A technology of fulvestrant and chemotherapeutic drugs, which is applied in the field of preparation of anti-microtubule chemotherapeutic drug resistance reversal agent of fulvestrant, achieving effects of less side effects, good clinical application prospects, and good patient tolerance
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Embodiment 1
[0021] Example 1: BCap37, Bats-72 and Bads-200P-glycoprotein and ER-α expression detection
[0022] (1) Experimental materials
[0023] Cell lines: paclitaxel-sensitive cell line BCap37 (purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences), two new cell lines Bats-72 and Bads-200 (strain numbers are CCTCC No: C201065 and CCTCC No: C201066, which have been in the prior application 201010502654.6 As a new cell line protection), the P-gp positive cell line MCF7 / ADR (purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences), and the estrogen receptor alpha positive cell line is MCF-7 (purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences);
[0024] Reagent consumables: 1640 culture medium (Hangzhou Jinuo Biomedical Technology Co., Ltd.), 0.25% trypsin (Hangzhou Jinuo), super neonatal bovine serum (Shanghai Yikesai Biological Products Co., Ltd.), PBS, Tween-20, 100mm culture Dishes (Corning), protein lysate (Beiyuntian), BCA ...
Embodiment 2
[0034] Example 2: Detection of drug resistance of Bats-72 and Bads-200 (taking paclitaxel as an example).
[0035] (1) Experimental materials
[0036] Cell lines: Paclitaxel-sensitive cell line BCap37, novel cell lines Bats-72 and Bads-200
[0037] Reagent consumables: 1640 culture medium (Hangzhou Gino), 0.25% trypsin (Hangzhou Gino), super neonatal bovine serum (Shanghai Ekesai), 100mm culture dish (Corning), 96-well plate (Corning), DMSO (Shanghai Sinopharm), MTT, Paclitaxel.
[0038] Instruments: cell incubator, ultra-clean bench, hemocytometer, microplate reader.
[0039] (2) Experimental method
[0040]BCap37 and Bads-200 were inoculated on 100mm culture dishes, cultivated for 72 hours, replaced with fresh culture medium, digested with 0.25% trypsin, collected cells, counted cells, prepared single cell suspension at a concentration of 20000 cells / ml, and inoculated 0.2ml per well The single cell suspension was transferred to a 96-well plate with 4000 cells per total,...
Embodiment 3
[0045] Example 3: Toxicity detection of fulvestrant on Bats-72 and Bads-200 cells.
[0046] (1) Experimental materials
[0047] Cell lines: new cell lines Bats-72 and Bads-200;
[0048] Reagent consumables: 1640 culture medium (Hangzhou Gino), 0.25% trypsin (Hangzhou Gino), super neonatal bovine serum (Shanghai Ekesai), 100mm culture dish (Corning), 96-well plate (Corning), DMSO (Shanghai Sinopharm), MTT, Paclitaxel.
[0049] Instruments: cell incubator, ultra-clean bench, hemocytometer, microplate reader.
[0050] (2) Experimental method
[0051] Bats-72 and Bads-200 were inoculated on 100mm culture dishes, cultured 72, replaced with fresh culture medium, digested with 0.25% trypsin, collected cells, counted cells, prepared a single cell suspension with a concentration of 20000 / ml, inoculated 0.2 ml single cell suspension to 96-well plate, each total cell number is 4000, cultivate overnight, add fulvestrant to treat the concentration gradient of 0, 10, 20, 50, 100, 200, 5...
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Abstract
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