Role of calcium in phototoxicity of 2-butylamino-2-demethoxy-hypocrellin A to human gastric cancer MGC-803 cells.
After incubation with 2-butylamino-2-demethoxy-hypocrellin A (2-BA-2-DMHA), photodynamically induced change in the cytoplasmic free calcium concentration ([Ca(2+)](i)) and its effect on cell damage were investigated in human gastric cancer (MGC-803). Fluorescence spectrophotometry measurement indicated that the photosensitization of MGC-803 by 2-BA-2-DMHA caused an increase in intracellular calcium [Ca(2+)](i), and this increase in [Ca(2+)](i) showed a dependence on the concentration of 2-BA-2-DMHA, light dose and extracellular [Ca(2+)](e). This phenomenon of intracellular calcium accumulation was further confirmed by using laser scanning confocal microscopy (LSCM). Furthermore, the results from MTT assay and flow cytometry analysis suggested that chelation of extracellular calcium by EGTA or intracellular calcium by BAPTA could inhibit photodynamically induced cell killing, while increase of [Ca(2+)](i) by thapsigargin (TG), a highly specific inhibitor of the Ca(2+)-ATPase, or by A23187, a calcium ionophore could enhance this action. Meanwhile, the nucleus morphology was also investigated by fluorescence microscopy. The results indicated that the increase in intracellular Ca(2+) concentration was responsible for 2-BA-2-DMHA photodynamically induced damage to MGC-803.
Zhou
Zhixiang
Z
Center for Molecular Biology, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, 100101, Beijing, PR China.
Yang
Hongying
H
Zhang
Zhiyi
Z
D000001:Calcimycin; D002118:Calcium / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D002614:Chelating Agents; D004533:Egtazic Acid / Q000031:analogs & derivatives*; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D016257:Fura-2; D006801:Humans; D007476:Ionophores; D008027:Light; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D010569:Perylene / Q000031:analogs & derivatives* / Q000494:pharmacology*; D010778:Photochemotherapy; D017319:Photosensitizing Agents / Q000494:pharmacology*; D011809:Quinones / Q000494:pharmacology*; D013274:Stomach Neoplasms / Q000188:drug therapy*; D019284:Thapsigargin; D014407:Tumor Cells, Cultured
C114700:2-butylamino-2-demethoxy-hypocrellin A; D002614:Chelating Agents; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D007476:Ionophores; D017319:Photosensitizing Agents; D011809:Quinones; D000001:Calcimycin; D004533:Egtazic Acid; D010569:Perylene; D019284:Thapsigargin; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D002118:Calcium; D016257:Fura-2