Department of Molecular Biology and Genetics, Faculty of Science, Istanbul University, Vezneciler, 34134, Istanbul, Turkey.
Cell Stress Chaperones. 2018 Sep;23(5):885-896. doi: 10.1007/s12192-018-0896-z. Epub 2018 Apr 7.
High expression of Hsp27 in glioma cells has been closely associated with tumor cell proliferation and apoptosis inhibition. The aim of the present study was to asses the effects of rosmarinic acid (RA) on Hsp27 expression and apoptosis in non-transfected and transfected human U-87 MG cells. The effect of rosmarinic acid was compared to quercetin, which is known to be a good Hsp27 inhibitor. In order to block the expression of Hsp27 gene (HSPB1), transfection with specific siRNAs was performed. Western blotting technique was used to assess the Hsp27 expression, and caspase-3 colorimetric activity assay was performed to determine apoptosis induction. According to the results, it was found that RA and quercetin effectively silenced Hsp27 and both agents induced apoptosis by activating the caspase-3 pathway. Eighty and 215 μM RA decreased the level of Hsp27 by 28.8 and 46.7% and induced apoptosis by 30 and 54%, respectively. For the first time, we reported that rosmarinic acid has the ability to trigger caspase-3 induced apoptosis in human glioma cells. As a result of siRNA transfection, the Hsp27 gene was silenced by ~ 50% but did not cause a statistically significant change in caspase-3 activation. It was also observed that apoptosis was induced at a higher level as a result of Hsp27 siRNA and subsequent quercetin or RA treatment. siRNA transfection and 215 μM RA treatment suppressed Hsp27 expression level by 90.5% and increased caspase-3 activity by 58%. Herein, we demonstrated that RA administered with siRNA seems to be a potent combination for glioblastoma therapy.
热休克蛋白 27（Hsp27）在神经胶质瘤细胞中的高表达与肿瘤细胞增殖和凋亡抑制密切相关。本研究旨在评估迷迭香酸（RA）对未转染和转染的人 U-87 MG 细胞中 Hsp27 表达和凋亡的影响。将迷迭香酸的作用与槲皮素进行了比较，槲皮素已知是一种良好的 Hsp27 抑制剂。为了阻断 Hsp27 基因（HSPB1）的表达，进行了特异性 siRNA 转染。采用 Western blot 技术评估 Hsp27 表达，采用 caspase-3 比色活性测定法检测细胞凋亡诱导。结果发现，RA 和槲皮素有效地沉默了 Hsp27，两种药物均通过激活 caspase-3 途径诱导细胞凋亡。80 和 215μM RA 使 Hsp27 水平分别降低了 28.8%和 46.7%，并分别诱导了 30%和 54%的凋亡。我们首次报道迷迭香酸具有触发人神经胶质瘤细胞 caspase-3 诱导凋亡的能力。由于 siRNA 转染，Hsp27 基因被沉默了约 50%，但对 caspase-3 激活没有引起统计学上的显著变化。还观察到由于 Hsp27 siRNA 及随后的槲皮素或 RA 处理，凋亡诱导水平更高。siRNA 转染和 215μM RA 处理使 Hsp27 表达水平降低了 90.5%，并使 caspase-3 活性增加了 58%。在此，我们证明了与 siRNA 联合使用的 RA 似乎是治疗神经胶质瘤的有效组合。
