Methylstat is a potent histone demethylases inhibitor. Methylstat shows anti-proliferative activity with low cytotoxicity. Methylstat induces Apoptosis and cell cycle arrest at G0/G1 phase. Methylstat increases the expression of p53 and p21 protein levels. Methylstat inhibits angiogenesis induced by various cytokines. Methylstat can be used as a chemical probe for addressing its role in angiogenesis[1][2].
Methylstat (0-5 µM; 48, 72 h) shows anti-proliferative activity with no cytotoxicity on HUVECs at 1-2 µM[1].
Methylstat (0, 1, 2 µM; 48 h) induces cell cycle arrest at G0/G1 phase in a dose-dependent manner[1].
Methylstat (0, 1, 2 µM; 48 h) increases the expression of p53 mRNA levels, the H3K27 methylation levels and the accumulation of p53 and p21 protein levels, but suppresses the protein level of cyclinD1[1].
Methylstat (0, 1, 2 µM) shows anti-angiogenic activity induced by VEGF, bFGF and TNF-α in HUVEC cells, and inhibits the f capillary formation during CAM (chick embryo chorioallantoic membrane) development without any sign of thrombosis and hemorrhage[1].
Methylstat (1.1, 2.2 mM for U266 cells, 2.1, 4.2 mM for ARH77 cells; 72 h) induces apoptosis significantly in U266 and ARH77 cells[2].
Cell Cytotoxicity Assay[1]
| Cell Line: | HUVEC cells |
| Concentration: | 0-5 µM |
| Incubation Time: | 48, 72 h |
| Result: | Did not exhibit cytotoxicity on HUVECs at 1-2 µM. |
Cell Viability Assay[1]
| Cell Line: | HUVEC, HepG2, HeLa, CHANG cells |
| Concentration: | 0-5 µM |
| Incubation Time: | 72 h |
| Result: | Showed anti-proliferative activity with IC50s of 4, 10, 5, 7.5 µM for HUVEC, HepG2, HeLa, CHANG cells, respectively. |
Cell Cycle Analysis[1]
| Cell Line: | HUVEC cells |
| Concentration: | 0, 1, 2 µM |
| Incubation Time: | 48 h |
| Result: | G0/G1 phase increased 16.8% compared to non-treated cells, whereas S andG2/M decreased 5.5% and 6.1% respectively. |
Western Blot Analysis[1]
| Cell Line: | HUVEC cells |
| Concentration: | 0, 1, 2 µM |
| Incubation Time: | 0-48 h |
| Result: | Resulted in accumulation of p53 and p21 protein levels in a time- and dose-dependent manner and increased the H3K27 methylation levels, the but suppressed the protein level of cyclinD1. |
Apoptosis Analysis[2]
| Cell Line: | U266, ARH77 cells |
| Concentration: | 1.1, 2.2 mM for U266 cells, 2.1, 4.2 mM for ARH77 cells |
| Incubation Time: | 72 h |
| Result: | Induced apoptosis in U266, ARH77 cells. |
[1]. Yumi Cho, et al. A histone demethylase inhibitor, methylstat, inhibits angiogenesis in vitro and in vivo. RSC Advances, 2014.
[2]. Kac? FN, et al. Synergistic Apoptotic Effects of Bortezomib and Methylstat on Multiple Myeloma Cells. Arch Med Res. 2020 Apr;51(3):187-193.