MCF-7 breast carcinoma cells express ryanodine receptor type 1: functional characterization and subcellular localization.
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.
Saldaña
Carlos
C
Cellular and Molecular Neurobiology Department, Neurobiology Institute, Campus UNAM-Juriquilla, 76230, Querétaro, QRO, Mexico.
Díaz-Muñoz
Mauricio
M
Antaramián
Anaid
A
González-Gallardo
Adriana
A
García-Solís
Pablo
P
Morales-Tlalpan
Verónica
V
D000368:Aged; D000818:Animals; D001483:Base Sequence; D001943:Breast Neoplasms / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D045744:Cell Line, Tumor; D004791:Enzyme Inhibitors / Q000378:metabolism; D005260:Female; D006801:Humans; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D019837:Ryanodine Receptor Calcium Release Channel / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D015398:Signal Transduction / Q000502:physiology; D013347:Subcellular Fractions / Q000378:metabolism; D019284:Thapsigargin / Q000378:metabolism
D004791:Enzyme Inhibitors; D053496:Inositol 1,4,5-Trisphosphate Receptors; D020033:Protein Isoforms; D019837:Ryanodine Receptor Calcium Release Channel; D019284:Thapsigargin; D002118:Calcium