Application of DTTZ in preparation of medicine for preventing and treating chemotherapy injury
A technology of drugs and chemotherapeutic agents, applied in the field of biomedicine, can solve problems such as prevention and treatment effects that have not been mentioned
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Embodiment 1
[0028] Embodiment 1 Neutral red test detects the cell selectivity and optimal concentration of DTTZ hydrochloride chemotherapy protection
[0029] Select normal cells including human intestinal epithelial cells (HIEC-6), hamster ovary cells (CHO) and cancer cells including human breast cancer (MCF-7), human non-small cell lung cancer (A549) and human cervical cancer (Hela) etc. 5 Seed cells to investigate the protective effect of DTTZ on normal cells of chemotherapy drugs cisplatin and doxorubicin. Cells were cultured in DMEM with 10% fetal bovine serum at 37°C, 5% CO2 Cultivated in an incubator. The cells were seeded in a 96-well plate at a density of 5,000 cells / well. After 24 h, the cells were incubated with DTTZ or the positive control WR2721 for 15 min in advance, and then chemotherapeutic drugs (cisplatin or doxorubicin) were added to continue the incubation. The wells were washed twice with PBS, and the neutrophil proliferation and cytotoxicity detection kit was used t...
Embodiment 2
[0031] Example 2 EdU staining to detect the protective effect of DTTZ hydrochloride chemotherapy on the DNA synthesis process of normal cells
[0032] EdU (5-ethynyl-2'-deoxyuridine), Chinese name is 5-ethynyl-2'-deoxyuridine, is a new type of thymidine (thymidine, thymidine) analog, EdU can be synthesized in DNA In the process, alternative thymidine is incorporated into newly synthesized DNA. If DNA synthesis is inhibited, EdU incorporation is also inhibited. Therefore, the inhibitory effect of the drug on cell DNA synthesis can be judged according to the number of cells stained with EdU. HIEC-6 and MCF-7 were seeded in 24-well plates at a density of 50,000 cells / well for 24 hours, then DTTZ or positive control WR2721 was incubated for 15 minutes in advance, and the chemotherapeutic drug cisplatin was added to continue incubation. After 24 hours, BeyoClickTM EdU-488 was used for detection Cells were treated with the kit and photographed under an inverted fluorescence micros...
Embodiment 3
[0034] Example 3 Clone formation experiment to investigate the cytotoxicity of DTTZ and positive control WR2721
[0035] Cell clone formation assay is an important technical method used to detect cell proliferation ability, invasiveness, population dependence, etc. Compared with cell survival assays (such as MTT assay), the clone formation rate reflected by the drug is more sensitive to the effect of cell proliferation. If the cell's genetic material is disturbed by drugs, it will not be possible to form a clonal population from a single cell through the process of mitosis. Therefore, clonogenic assays can be used to investigate the toxicity of drugs to cells.
[0036] Four kinds of normal cells including human intestinal epithelial cells (HIEC-6), human renal epithelial cells (293T) and cancer cells including human breast cancer (MCF-7) and human cervical cancer (Hela) were selected to investigate DTTZ and the positive control WR2721 cytotoxicity. The cells were resuspende...
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