Laboratório de Farmacologia Molecular, Universidade de Brasília, Av. L2 Norte, Brasília, DF 70.910-900, Brazil; Laboratório de Hematologia e Células-tronco, Universidade de Brasília, Av. L2 Norte, Brasília, DF 70.910-900, Brazil.
Laboratório de Hematologia e Células-tronco, Universidade de Brasília, Av. L2 Norte, Brasília, DF 70.910-900, Brazil.
Toxicol In Vitro. 2020 Dec;69:104992. doi: 10.1016/j.tiv.2020.104992. Epub 2020 Sep 2.
Euchromatic histone-lysine N-methyltransferase 1 (EHMT1) and EHMT2 are upregulated in various human cancers, and their deregulation is associated with tumor development and progression. In this paper, we investigated the expression level of EHMT1/EHMT2 in acute lymphoblastic leukemia (ALL) and whether the modulation of these enzymes could have any cellular or molecular impact on ALL cells. For this, we used UNC0646 as a priming strategy to target EHMT1/EHMT2 and investigated its effect on proliferation and cell viability of Jurkat cells by MTT assay. Then, considering the IC50 and IC75, cellular death was determined by Annexin V/PI staining using flow cytometry. Finally, we investigated by RT-PCR the molecular bases that could be involved in the observed effects. Interestingly, accessing the International Microarray Innovations in Leukemia (MILE) study group, we detected that both EHMT1 and EHMT2 are overexpressed in ALL. More important, we determined that inhibition of EHMT1/EHMT2 significantly decreased Jurkat cell viability in a dose-dependent manner. Accordingly, we observed that inhibition of EHMT1/EHMT2 promoted Jurkat cell death, which was accompanied by increased expression of P53, TP73, BAX, and MDM4. These results clearly indicate that inhibition of EHMT1/EHMT2 induces pro-apoptotic gene expression in ALL and promotes cell death. More importantly, the modulation of these histone methyltransferases may be a promising epigenetic target for ALL treatment.
euchromatic histone-lysine N-methyltransferase 1 (EHMT1) 和 EHMT2 在各种人类癌症中上调，其失调与肿瘤的发生和发展有关。在本文中，我们研究了 EHMT1/EHMT2 在急性淋巴细胞白血病 (ALL) 中的表达水平，以及这些酶的调节是否对 ALL 细胞具有任何细胞或分子影响。为此，我们使用 UNC0646 作为靶向 EHMT1/EHMT2 的启动策略，并通过 MTT 测定法研究其对 Jurkat 细胞增殖和细胞活力的影响。然后，考虑到 IC50 和 IC75，通过流式细胞术用 Annexin V/PI 染色来确定细胞死亡。最后，我们通过 RT-PCR 研究了可能参与观察到的效应的分子基础。有趣的是，我们访问了国际白血病微阵列创新 (MILE) 研究组，发现 EHMT1 和 EHMT2 在 ALL 中均过表达。更重要的是，我们确定抑制 EHMT1/EHMT2 以剂量依赖性方式显著降低 Jurkat 细胞活力。相应地，我们观察到抑制 EHMT1/EHMT2 促进 Jurkat 细胞死亡，这伴随着 P53、TP73、BAX 和 MDM4 的表达增加。这些结果清楚地表明，抑制 EHMT1/EHMT2 在 ALL 中诱导促凋亡基因表达并促进细胞死亡。更重要的是，这些组蛋白甲基转移酶的调节可能是 ALL 治疗的有前途的表观遗传靶点。
