Inhibition of phosphodiestrase 9 induces cGMP accumulation and apoptosis in human breast cancer cell lines, MCF-7 and MDA-MB-468
Inhibition of phosphodiestrase 9 induces cGMP accumulation and apoptosis in human breast cancer cell lines, MCF-7 and MDA-MB-468
复制标题
DOI：
10.1111/j.1365-2184.2012.00819.x
复制
发表时间：
2012-06-01
影响因子：
8.5
通讯作者：
Edalat, R.
中科院分区：
文献类型：
--
作者：
Saravani, R.;Karami-Tehrani, F.;Edalat, R.
目的：磷酸二酯酶9（PDE 9）是磷酸二酯酶水解cGMP的主要亚型，并且通过细胞内cGMP信号传导在细胞增殖、细胞分化和细胞凋亡中起关键作用。本研究旨在研究PDE 9对人乳腺癌细胞系MCF- 7和MDA-MB- 468的表达、活性和凋亡作用。材料和方法：分别采用比色法和真实的RT-PCR法检测PDE 9的活性和表达，并测定cGMP浓度。MTT法、Annexin V-FITC染色、Hoechst 33258染色和caspase 3活性测定检测细胞凋亡。结果如下：用BAY 736691处理两种细胞系导致PDE 9 mRNA表达、PDE 9 cGMP水解活性降低和细胞内cGMP应答升高。BAY 73- 6691以剂量和时间依赖性方式显著降低细胞增殖，并通过激活caspase 3引起凋亡显著增加。结论：BAY 736691通过cGMP途径诱导乳腺癌细胞凋亡。这些数据表明BAY 73- 6691可用作治疗乳腺癌的药剂。
Objectives: Phosphodiesterase 9 ( PDE9) is a major isoform of phosphodiesterase hydrolysing cGMP and plays a key role in proliferation of cells, their differentiation and apoptosis, via intracellular cGMP signalling. The study described here was designed to investigate expression, activity and apoptotic effect of PDE9 on human breast cancer cell lines, MCF- 7 and MDA- MB- 468. Materials and methods: Activity and expression of PDE9 were examined using colorimetric cyclic nucleotide phosphodiesterase assay and real- time RT- PCR methods respectively; cGMP concentration was also measured. MTT viability test, annexin V- FITC staining, Hoechst 33258 staining and caspase3 activity assay were used to detect apoptosis. Results: Treatment of both cell lines with BAY 736691 lead to reduction in PDE9 mRNA expression, PDE9 cGMP- hydrolytic activity and elevation of the intracellular cGMP response. BAY 73- 6691 significantly reduced cell proliferation in a doseand time- dependent manner and caused marked increase in apoptosis through caspase3 activation. Conclusion: Our results revealed that BAY 736691 induced apoptosis in these breast cancer cell lines through the cGMP pathway. These data suggest that BAY 73- 6691 could be utilized as an agent in treatment of breast cancer.