Loss of mitochondrial membrane potential is inhibited by bombesin in etoposide-induced apoptosis in PC-3 prostate carcinoma cells
Loss of mitochondrial membrane potential is inhibited by bombesin in etoposide-induced apoptosis in PC-3 prostate carcinoma cells
复制标题
DOI：
10.1158/1535-7163.mct-06-0681
复制
发表时间：
2007-04-01
影响因子：
5.7
通讯作者：
Vilches, Jose
中科院分区：
文献类型：
--
作者：
Salido, Mercedes;Gonzalez, Juan L.;Vilches, Jose
神经内分泌分泌产物及其与前列腺上皮细胞的相互作用目前正在研究中，以了解其在前列腺癌的发病机制，预后和治疗中的意义。这些神经肽有可能破坏肿瘤中细胞死亡和细胞生长之间的平衡。我们的研究是基于蛙皮素在调节线粒体膜电位[Δ psi（m））在依托泊苷诱导的PC-3细胞死亡中的作用。培养细胞并用5，5 ′，6，6 ′-四氯-1，1 ′，3，3 ′-四乙基苯并咪唑基羰花青碘化物（JC-1）染色。在低膜电位下，JC-1产生绿色荧光，在高膜电位下，它形成具有红色荧光的"J聚集体”。在共聚焦显微镜下检查细胞。对于定量分析，选择感兴趣的区域。计算每个感兴趣区域中红色和绿色通道中的大小、像素数和荧光强度之间的比率。蛙皮素可阻止依托泊苷处理的PC-3细胞中Δ psi（m）的损失。JC-1染色细胞的定量分析显示，与对照细胞相比，依托泊苷处理的细胞中红色（高Δ psi（m））与绿色（低Δ psi（m））的比率显著降低，在蛙皮素存在下恢复（P < 0.00001）。处理和面积之间的相互作用[P = 0.0002）是高度显著的，并且证实PC-3细胞保持其凋亡机制，显示响应于依托泊苷的凋亡体积减小。蛙皮素的保护作用是通过抑制细胞凋亡和维持线粒体的完整性来实现的。需要开发新的治疗方案和试验来测试通过中和神经内分泌激素介导的抗凋亡细胞内途径起作用的药物。
Neuroendocrine secretory products and their interactions with epithelial prostate cells are currently under investigation in order to understand their significance in the pathogenesis, prognosis, and therapy of prostate carcinoma. These neuropeptides have the potential to disrupt the balance between cell death and cell growth in the tumor. Our research was based on the role of bombesin in modulating the mitochondrial membrane potential [Delta psi(m)) in cell death induced by etoposide on PC-3 cells. Cells were cultured and stained with 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide (JC-1). At low membrane potentials, JC-1 produces a green fluorescence, and at high membrane potentials, it forms '' J aggregates" with red fluorescence. Cells were examined in a confocal microscope. For quantitative analyses, regions of interest were selected. The size, number of pixels, and ratios between fluorescence intensity in the red and green channels in each region of interest were calculated. The loss of Delta psi(m) in etoposide-treated PC-3 cells was prevented by bombesin. The quantitative analysis of JC-1-stained cells revealed a significant decrease in the red (high Delta psi(m)) to green (low Delta psi(m)) ratio in etoposide-treated cells when compared with control cells, which was restored in the presence of bombesin (P < 0.00001). The interaction between treatments and area [P = 0.0002) was highly significant, and confirms that PC-3 cells keep their apoptosis machinery, showing an apoptotic volume decrease in response to etoposide. The protection by bombesin occurs by inhibition of apoptosis and maintenance of mitochondrial integrity. New therapeutic protocols and trials need to be developed to test drugs acting through the neutralization of antiapoptotic intracellular pathways mediated by neuroendocrine hormones.