Saldaña Carlos, Díaz-Muñoz Mauricio, Antaramián Anaid, González-Gallardo Adriana, García-Solís Pablo, Morales-Tlalpan Verónica
Cellular and Molecular Neurobiology Department, Neurobiology Institute, Campus UNAM-Juriquilla, 76230, Querétaro, QRO, Mexico.
Mol Cell Biochem. 2009 Mar;323(1-2):39-47. doi: 10.1007/s11010-008-9962-7. Epub 2008 Dec 4.
Breast carcinoma-derived MCF-7 cells are frequently used in biomedical research. However, few reports exist regarding the characterization of signaling mechanisms in these cancerous cells involved in intracellular Ca(2+) dynamics. Consequently, the aim of these experiments was to characterize the ryanodine receptor/Ca(2+) release channel (RyR) present in MCF-7 cells. Ryanodine (100 nM), cADPR (5 microM), and caffeine (10 mM) promoted cytoplasmic Ca(2+) mobilization; in contrast, ryanodine at inhibitory concentration (100 microM) decreased the basal Ca(2+) level. Fluorescent probes demonstrated that RyR is located mainly in endomembranes. Some degree of co-localization with inositol trisphosphate receptor (IP(3)R) was observed, whereas coincidence with thapsigargin-sensitive Ca(2+)-ATPase (SERCA) was more limited. Molecular cloning resulted in the detection exclusively of RyR isoform 1. For the first time, it is shown that MCF-7 cells express functional RyR.
源自乳腺癌的MCF-7细胞常用于生物医学研究。然而，关于这些癌细胞中参与细胞内Ca(2+)动态变化的信号传导机制的特征报道却很少。因此，这些实验的目的是表征MCF-7细胞中存在的兰尼碱受体/Ca(2+)释放通道（RyR）。兰尼碱（100 nM）、环ADP核糖（cADPR，5 microM）和咖啡因（10 mM）促进细胞质Ca(2+)动员；相反，抑制浓度的兰尼碱（100 microM）降低基础Ca(2+)水平。荧光探针表明RyR主要位于内膜。观察到与肌醇三磷酸受体（IP(3)R）有一定程度的共定位，而与毒胡萝卜素敏感的Ca(2+)-ATP酶（SERCA）的共定位则较为有限。分子克隆仅检测到RyR亚型1。首次表明MCF-7细胞表达功能性RyR。
