Department of Molecular Cell Biology, Institute of Biochemistry, Vilnius University, 08662 Vilnius, Lithuania.
Mol Biol Rep. 2012 Dec;39(12):10179-86. doi: 10.1007/s11033-012-1892-5. Epub 2012 Sep 26.
Histone deacetylase inhibitors (HDACIs) represent a new class of targeted anti-cancer agents and different other diseases, like muscular disorders. A number of studies have shown that extracellular signal-activated kinases can target chromatin-modifying complexes directly and regulate their function. The molecular connection between the dystrophin-associated protein complex (DAPC) and chromatin has been described, by showing that NO signaling regulates histone deacetylase (HDAC) activity and influences gene expression in different cell types. In present study, we investigated HDACs changes in HeLa cells undergoing growth inhibition and apoptosis, caused by HDACI BML-210 and retinoic acid (ATRA). Cell cycle analysis indicated that HeLa cell treatment with 20 and 30 μM concentration of BML-210 increased the proportion of cells in G0/G1 phase, and caused accumulation in subG1, indicating that the cells are undergoing apoptosis. We determined down-regulation of HDAC 1-5 and 7 after treatment with BML-210. Also, we demonstrated expression of different isoforms of alpha-dystrobrevin (α-DB) and other components of DAPC such as syntrophin, dystrophin, beta-dystrobrevin (β-DB) and NOS in HeLa cells after treatments. We determined changes in protein expression level of dystrophin, NOS1, α- and β-DB and in subcellular localization of α-DB after treatments with BML-210 and ATRA. In conclusion, these results suggest that HDACI BML-210 can inhibit cell growth and induce apoptosis in cervical cancer cells, what correlates with down-regulation of HDAC class I and II and changes in the DAPC expression levels. This can be important for identifying target proteins in DAPC signaling to HDACs, as a target of pharmacological intervention for treatment of muscular dystrophies and other diseases.
组蛋白去乙酰化酶抑制剂 (HDACIs) 代表了一类新的靶向抗癌药物和其他疾病的药物,如肌肉疾病。许多研究表明,细胞外信号激活的激酶可以直接靶向染色质修饰复合物并调节其功能。已经描述了与营养不良相关蛋白复合物 (DAPC) 和染色质之间的分子连接,表明 NO 信号调节组蛋白去乙酰化酶 (HDAC) 活性并影响不同细胞类型的基因表达。在本研究中,我们研究了 HeLa 细胞中 HDACs 的变化,这些细胞受到 HDACI BML-210 和维甲酸 (ATRA) 的生长抑制和凋亡的影响。细胞周期分析表明,用 20 和 30 μM 浓度的 BML-210 处理 HeLa 细胞会增加 G0/G1 期细胞的比例,并导致亚 G1 期积累,表明细胞正在凋亡。我们确定在用 BML-210 处理后 HDAC1-5 和 7 的下调。此外,我们还证明了在 HeLa 细胞中表达不同的α-肌营养不良蛋白 (α-DB) 同工型和 DAPC 的其他成分,如肌联蛋白、肌营养不良蛋白、β-肌营养不良蛋白 (β-DB) 和 NOS。我们确定了在用 BML-210 和 ATRA 处理后,α-DB 的蛋白表达水平和亚细胞定位的变化。总之,这些结果表明,HDACI BML-210 可以抑制宫颈癌细胞的生长并诱导细胞凋亡,这与 HDAC 类 I 和 II 的下调以及 DAPC 表达水平的变化相关。这对于鉴定 DAPC 信号传导到 HDACs 的靶蛋白很重要,作为治疗肌肉疾病和其他疾病的药理学干预的靶标。