| Ca2+-dependent and Caspase-3–independent Apoptosis Caused by Damage in Golgi Apparatus due to 2,4,5,7-Tetrabromorhodamine 123 Bromide–induced Photodynamic Effects¶ | |
Abstract/OtherAbstract
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To clarify the role of the Golgi apparatus in photodynamic therapy–induced apoptosis, its signaling pathway was studied after photodynamic treatment of human cervix carcinoma cell line HeLa, in which a photosensitizer, 2,4,5,7-tetrabromorhodamine 123 bromide (TBR), was incorporated into the Golgi apparatus. Laser scanning microscopic analysis of TBR-loaded HeLa cells confirmed that TBR was exclusively located in the Golgi apparatus. HeLa cells incubated with TBR for 1 h were then exposed to visible light using an Xe lamp. Light of wavelength below 670 nm was eliminated with a filter. Morphological observation of nuclei stained with Hoechst 33342 revealed that apoptosis of cells was induced by exposure to light. Electron spin resonance spectrometry showed that light-exposed TBR produced both singlet oxygen (1O2) and superoxide anion (O2–). Apoptosis induction by TBR was inhibited by pyrrolidine dithiocarbamate, an O2– scavenger, but not by NaN3, a quencher of 1O2. Furthermore, TBR-induced apoptosis was inhibited by aurintricarboxylic acid and ZnCl2, which are known as inhibitors of deoxyribonuclease (DNase) γ, and (acetoxymethyl)-1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid, a chelator of Ca2+, but not by acetyl Asp-Glu-Val-Asp-aldehyde, an inhibitor of caspase-3. These results suggested that O2– was responsible for TBR-induced apoptosis, and Ca2+-dependent and caspase-3–independent nuclease such as DNase γ played an important role in apoptotic signaling triggered by Golgi dysfunction. |
Authors
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Maiko Ogata, Osamu Inanami, Mihoko Nakajima, Takayuki Nakajima, Wakako Hiraoka, Mikinori Kuwabara |
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Publisher : American Society for Photobiology Type : Text Format : text/html |
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2003-09 |
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Languages : en-US |
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