Saravani Ramin, Galavi Hamid Reza, Shahraki Ali
Cellular and Molecular Research Center, Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran.
Department of Biology, Faculty of Science, University of Sistan and Baluchestan, Zahedan, Iran.
Breast Cancer (Auckl). 2017 Feb 23;11:1178223417690178. doi: 10.1177/1178223417690178. eCollection 2017.
This study aimed to investigate the effect of hydroalcoholic C. Koch extract (HAWE) on phosphodiesterase 5 (PDE5) gene expression and cyclic guanosine 3',5' monophosphate (cGMP) signaling in the MCF-7 and MDA-Mb-468 cell lines. The effective dose (ED50) of HAWE was examined in both cell lines using a 3-(4,5-dimethylhiazol-2-yl)-2,5-diphenyltetrazolium bromide viability test, and the type of cell death was detected by flow cytometry. The expression of PDE5 and the concentration of cGMP were measured in a time-dependent manner in the ED50 by real-time polymerase chain reaction and a colorimetric assay, respectively. Treatment with HAWE showed 25 µg/mL to be the ED50 for both cell lines, and HAWE led to a reduction in the PDE5 messenger RNA expression. The intracellular cGMP increased in a time-dependent manner. The results showed that HAWE has an antiproliferative property in MCF-7 and MDA-Mb-468 cell lines through the cGMP pathway. Therefore, HAWE is a potential source to effectively isolate inhibitory PDE5.
本研究旨在探讨水醇提取物柯氏梭菌(HAWE)对MCF-7和MDA-Mb-468细胞系中磷酸二酯酶5(PDE5)基因表达及环磷酸鸟苷(cGMP)信号传导的影响。使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐活力试验在两种细胞系中检测HAWE的有效剂量(ED50),并通过流式细胞术检测细胞死亡类型。分别通过实时聚合酶链反应和比色法以时间依赖性方式在ED50中测量PDE5的表达和cGMP的浓度。用HAWE处理显示两种细胞系的ED50均为25 µg/mL,且HAWE导致PDE5信使核糖核酸表达降低。细胞内cGMP以时间依赖性方式增加。结果表明,HAWE通过cGMP途径在MCF-7和MDA-Mb-468细胞系中具有抗增殖特性。因此,HAWE是有效分离抑制性PDE5的潜在来源。