DNA methyltransferase enzymes (DNMTs) are responsible for the DNA methylation process which could catalyze the transfer of a methyl group from S-adenosyl methionine to the cytosine target nucleotide producing methylcytosine[5].
体内研究
The nude mice with SKOV3 xenografts were treated with 2 mg/kg 5-AZ thrice weekly for 10 weeks. The volume and weight per nodule were decreased after the treatment of 5-AZ, which indicated that the 5-AZ could inhibit growth of tumors[8].
体外研究
Azacytidine (5-AZ) is a DNMT inhibitor with IC50 value of 0.8 - 3 μmol/L on MM cells[6]. The K12-72.1 cells were treated with 5-AZ at both 5 and 10 μM, and the methylation status of the CpG sites in the promotor region of the gene PGP9.5 were measured by bisulfite sequencing. It was found that both concentrations could cause demethylation of the CpG sites and increased the expression of the gene PGP9.5. Moreover, after the treatment of 50 μM of 5-AZ on MCF10CA1a and MCF7 cells, the cell viability as measured by MTT assay decreased significantly compared with control[7].
作用机制
The 5-AZ could inhibit the DNA methylation through covalently binding to DNA methyltransferases, forming nucleoprotein adducts. Therefore, the number of active DNA methyltransferase enzymes in the cells are depletes[9].
5-Azacytidine/5-氮胞苷 细胞实验
Cell Line
Concentration
Treated Time
Description
References
4T1 cells
5 nM
48 hours
AZA+atRA reprogramming inhibited the growth of 4T1 cells
Cell Rep. 2023 Jun 27;42(6):112560.
T-HEp3 cells
5 nM
48 hours
AZA+atRA reprogramming inhibited the growth of T-HEp3 cells
Cell Rep. 2023 Jun 27;42(6):112560.
ERC cells
10 µM
24 h
Induce differentiation of ERC cells into cardiomyocytes
J Transl Med. 2007 Nov 15;5:57.
LLC1 cells
100nM
72 h
To evaluate the effect of 5-Azacytidine on the proliferation of LLC1 cells, results showed that 100nM 5-Azacytidine had limited effect on cell proliferation.
Nature. 2020 Mar;579(7798):284-290.
HNM007 cells
100nM
72 h
To evaluate the effect of 5-Azacytidine on the proliferation of HNM007 cells, results showed that 100nM 5-Azacytidine had limited effect on cell proliferation.
Nature. 2020 Mar;579(7798):284-290.
4T1 cells
100nM
72 h
To evaluate the effect of 5-Azacytidine on the proliferation of 4T1 cells, results showed that 100nM 5-Azacytidine had limited effect on cell proliferation.
Nature. 2020 Mar;579(7798):284-290.
Primary murine kidney fibroblasts
100 or 150 μg/ml
1 or 2 days
5′-Azacytidine effectively de-methylated Rasal1 and rescued endogenous Rasal1 expression, normalizing the proliferative activity of fibrotic renal fibroblasts.
EBioMedicine. 2015 Jan;2(1):19-36.
human cardiac stem cells (hCSCs)
10 μM
9 days
5-Azacytidine inhibits DNA methylation, leading to decreased levels of DNMT3a and SIRT1, thereby affecting epigenetic modifications and autophagy
Stem Cells. 2021 Apr;39(4):497-506.
THP-1 cells
10 μM
24 h
5-Azacytidine effectively blocked the LPS-induced upregulation of DNMT3b, increased PPARγ expression, and reduced IL-1β expression.
Adv Sci (Weinh). 2024 Nov;11(44):e2401931.
NSCLC cell lines
500 µM
24 h daily for 11 days
To investigate the effect of 5-Azacytidine in combination with HDAC inhibitors (e.g., ITF-2357) on the proliferation of NSCLC cells, results showed that the combination significantly inhibited cell proliferation.
Cell. 2017 Nov 30;171(6):1284-1300.e21.
A549 and H460 cells
500 µM
11 days
To investigate the inhibitory effect of 5-Azacytidine in combination with ITF-2357 on the proliferation of A549 and H460 cells, results showed that the combination significantly inhibited cell proliferation.
Cell. 2017 Nov 30;171(6):1284-1300.e21.
medaka fibroblast cells
2 μM
5 days
5-Azacytidine was used to reactivate the expression of Teratorn genes. The results showed that 5-azacytidine treatment moderately increased the expression of some Teratorn genes, although the expression level of most genes remained low.
Nat Commun. 2017 Sep 15;8(1):551.
OCI-LY1
0.3 µM
3 days
5-Azacytidine enhanced the sensitivity of OCI-LY1 cells to cisplatin, but the effect was less significant compared to other cell lines.
Clin Epigenetics. 2023 May 3;15(1):75.
SU-DHL2
0.3 µM
3 days
5-Azacytidine significantly enhanced the sensitivity of SU-DHL2 cells to cisplatin, reducing the IC50 value below the clinically achievable concentration.
Clin Epigenetics. 2023 May 3;15(1):75.
SU-DHL8
0.3 µM
3 days
5-Azacytidine significantly enhanced the sensitivity of SU-DHL8 cells to cisplatin, reducing the IC50 value below the clinically achievable concentration.
Clin Epigenetics. 2023 May 3;15(1):75.
5-Azacytidine/5-氮胞苷 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Mice
Balb/c mice
Intraperitoneal injection
5 mg/kg
Following the specified schedule
Inhibit DNMT1 to reduce CTX-induced myeloid cell expansion
Front Immunol. 2021 Jan 25;11:594540
Mice
LLC and HNM007 pulmonary metastasis models
Subcutaneously injected
0.5mg/kg
Daily for 14 days
To evaluate the effect of 5-Azacytidine on pulmonary metastasis models, results showed that low dose 5-Azacytidine inhibited the accumulation of MDSCs in the lung and prolonged disease-free and overall survival.
Nature. 2020 Mar;579(7798):284-290.
Mice
Unilateral ureteral obstruction (UUO) model
Intraperitoneal injection
10 mg/kg
Every other day until the indicated time points
5′-Azacytidine significantly ameliorated renal fibrosis in mice, reduced fibrotic areas and fibroblast accumulation, and restored Rasal1 expression.
EBioMedicine. 2015 Jan;2(1):19-36.
Mice
LPS-induced acute lung injury model
Intraperitoneal injection
1 mg/kg
Once daily for 3 days
5-Azacytidine significantly alleviated LPS-induced lung tissue damage and alveolar-capillary barrier injury, and reduced inflammatory cytokine levels in BAL fluid.
Adv Sci (Weinh). 2024 Nov;11(44):e2401931.
Mice
NOD/SCID mouse model
Intraperitoneal injection
0.5 mg/kg
Every 14 days for a total of 28 days
To investigate the anti-tumor effect of 5-Azacytidine in combination with ITF-2357 in a NSCLC mouse model, results showed that the combination significantly reduced tumor volume and weight.
Cell. 2017 Nov 30;171(6):1284-1300.e21.
Nude mice
OCI-LY1 and SU-DHL2 xenograft models
Intraperitoneal injection
0.5 mg/kg
Once daily for 5 consecutive days
5-Azacytidine significantly enhanced the cytotoxic effect of cisplatin in the SU-DHL8 xenograft model, but the effect was not significant in the OCI-LY1 model.
搜索
