Method for preparing genitourinary system tumor cell ferroptosis model based on ferric ammonium citrate and application in preparation of antitumor drugs
A ferric ammonium citrate, urinary system technology, applied in the field of preparation of anti-tumor drugs, can solve the problems of low model stability and high toxicity, and achieve the effect of promoting ferroptosis and increasing expression
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[0020] The invention provides a method for preparing a ferroptosis model of genitourinary system tumor cells based on ferric ammonium citrate, comprising the following steps: when the density of cultured tumor cells reaches 65-75%, adding ferric ammonium citrate and continuing to cultivate for 20-30 hours , to get the ferroptosis model. Ammonium ferric citrate, also known as ammonium ferric citrate (AFC), 2-hydroxy-1,2,3-propane tricarboxylic acid ammonium iron, is a double salt of ferric citrate and ammonium citrate ( figure 1 ). AFC is usually added to dairy products, wheat flour for bread, biscuits, milk powder, etc., and is used in nutritional supplements (iron fortifiers), and is a very safe chemical substance. That is, AFC is a generally recognized safe food additive grade iron supplement (for the treatment of iron deficiency anemia), which is edible, has a high safety factor, and has little toxic and side effects, while many existing chemotherapeutic drugs are very tox...
Embodiment 1
[0033] 1. Cell culture:
[0034] Human genitourinary system tumor cell lines including breast cancer cell MCF7, ovarian cancer cell SK-OV-3, prostate cancer cell 22Rv1, and kidney cancer cell A-498 were purchased from the Cell Resource Center of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. The above cells were cultured in McCOY's 5A (SIGMA, M4892) medium containing NaHCO 3 2.2g / L, high-quality fetal bovine serum 10%. The culture environment is gas phase: air, 95%; carbon dioxide, 5%. Temperature: 37°C.
[0035] 2.MTT experiment:
[0036] Ferric ammonium citrate was purchased from SIGMA. It was made up to a storage concentration of 100 mg / ml in sterile PBS and sterilized by suction through a filter with a pore size of 0.22 μm. Take 2×10 3 / ml Tumor cells in good growth state were seeded in 96-well cell culture plates. On the next day, AFC was added so that the concentrations were 0.001, 0.01, 0.1, and 1.0 mg / ml respectively. Three parallel c...
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