| 生物活性： |
靶点：DNA Methyltransferase;Autophagy
通路：Epigenetics;Autophagy
背景说明：5-Azacytidine是一种胞苷核苷类似物，也是一种DNA 甲基化抑制剂，通过与 DNA 结合共价捕获 DNA甲基转移酶 (DNA methyltransferases)特异性抑制DNA甲基化。5-Azacytidine 通过降低 DNA 甲基化水平调节基因表达。
生物活性：5-Azacytidine (Azacitidine; 5-AzaC; Ladakamycin) is a nucleoside analogue of cytidine that specifically inhibits DNA methylation. 5-Azacytidine is incorporated into DNA to covalently trap DNA methyltransferases and contributes to reverse epigenetic changes. 5-Azacytidine induces cell autophagy.[1-4]
In Vitro：5-Azacytidine was first synthesized almost 40 years ago. It was demonstrated to have a wide range of anti-metabolic activities when tested against cultured cancer cells and to be an effective chemotherapeutic agent for acute myelogenous leukemia.The finding that 5-azacytidine was incorporated into DNA and that， when present in DNA， it inhibited DNA methylation， led to widespread use of 5-azacytidine and 5-aza-2-deoxycytidine (Decitabine) to demonstrate the correlation between loss of methylation in specific gene regions and activation of the associated genes. [1]
细胞实验：Twenty mL of cells (circa 1×104 cells/mL) are pipetted into sterilized culture tubes with screw caps and incubated at 37°C overnight. The experiment is initiated by the addition of 1 mL of 5-Azacytidine (5-azaCR) or medium for a given period (from 0 to 240 min) prior to the addition of 1 mL of metabolite (or medium). Cell growth is determined twice a day for 3 days by means of a Model A Coulter counter. To determine IDSO and ID90 values， 5 mL of L1210 cells (5×103 cells/mL) are incubated with the drug at 37°C for 3 days， and cell growth is determined.[3]
动物实验：For the in vivo experiments， leukemic mice (bearing circa 1×103 cells/animal) are given injections i.p. with 0.2 mL of 5-Azacytidine (5-azaCR) of a given concentration. Two hr later， the reaction is started by injecting 0.5 mL of labeled metabolite (TdR-3H or UR-3H， 10 /μCi/12.5 μg). After 1 hr， animals (3 mice/group) are killed by cervical fracture.[3]
激酶实验：A crude cell-free extract is isolated from LI 210 cells in culture by suspension of the cells in a given volume of 0.05mol/LTris-HCl buffer， pH 7.4， and sonic extraction with a Biosonik at 70% maximal output for 30 sec. The supernatant is collected after centrifugation at 105，000 × g for 60 min (4°C) in a Model L Spinco ultracentrifuge. The final protein concentration of the cell-free extracts is approximately 3 mg/mL. The extracts are used as the source of enzymes. Ribonucleotide reductase activity is measured. A unit of enzyme is defined as the amount that catalyzed dCMP synthesis at a rate of 1 mμmole/hr. The assay systems for the measurement of pyrimidine nucleoside (CR) and deoxynucleoside (TdR， CdR) kinases are essentially those described by Chu and Fischer. However， reactions are terminated by heating for 2 min in a boiling water bath， and the phosphorylated derivatives are isolated according to the method of Bach. Fifty-jul aliquots are applied to 1-inch discs of diethylaminoethyl paper， which are then placed in counting vials and eluted with 0.5 mL of 0.5 mol/LPCA. After 1 hr， 12 mL of Diotol are added， and the radioactivity is determined.[3]
数据来源文献：[1]. Christman JK. 5-Azacytidine and 5-aza-2-deoxycytidine as inhibitors of DNA methylation: mechanistic studies and their implications for cancer therapy. Oncogene. 2002 Aug 12;21(35):5483-95.
[2]. Creusot F， et al. Inhibition of DNA methyltransferase and induction of Friend erythroleukemia cell differentiation by 5-azacytidineand 5-aza-2-deoxycytidine. J Biol Chem. 1982 Feb 25;257(4):2041-8.
[3]. Li LH，et al. Cytotoxicity and mode of action of 5-azacytidine on L1210 leukemia. Cancer Res. 1970 Nov;30(11):2760-9.
[4]. Marycz K， et al. 5-Azacytidine?and?Resveratrol?Enhance?Chondrogenic?Differentiation?of?Metabolic?Syndrome-Derived?Mesenchymal?Stem?Cells?by?Modulating?Autophagy.Oxid Med Cell Longev.?2019 May 12;2019:1523140.
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| 基本信息： |
CAS：320-67-2
中文名称：阿扎胞苷(10mM in DMSO,无菌）
英文名称：5-Azacytidine(10mM in DMSO,Sterile）
分子式：C8H12N4O5
分子量：244.2
规格：1ml ; 1.5ml ; 0.5ml ; 0.3ml ; 0.1ml
溶解性：请根据自己的实验要求使用。
外观(性状)：无菌溶液
储存条件：Stroe at -20℃，6 months.
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| 靶点： |
DNA Methyltransferase;Autophagy |
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