吖啶橙經藍光照射後產生光毒性阻殺人類膀胱癌細胞
林宜佳1、林致凡2、蔡德甫1,4、陳宏恩1、仇光宇1,4、黃一勝1,3,4
1新光醫院外科部泌尿科、2新光醫院中央研究室、3台北醫學大學醫學院、4輔仁大學醫學院
Acridine orange exhibits photodamages in human bladder cancer cells under blue light exposure
Yi-Chia Lin1, Ji-Fan Lin2, Te-Fu Tsai 1,4,Hung-En Chen1, and Kuang-Yu Chou1,4, Thomas I-Sheng Hwang1,3,4
1Department of Urology, Shin Kong Wu Ho-Su Memorial Hospital ,2Central Laboratory, Shin Kong Wu Ho-Su Memorial Hospital Department of Surgery, Shin Kong Wu Ho-SuMemorial Hospital, 3Department of Urology, Taipei Medical University, Division of Urology, 4School of Medicine, Fu-Jen Catholic University, Taipei, Taiwan.
Background: Human bladder cancer (BC) cells exhibited a high basal level of autophagic activity demonstrated by accumulating of acridine orange (AO)-stained acidic vesicular organelles. In this study, we aim to investigate the cytotoxicity effects of AO on the human BC cell lines under blue-light exposure.
Materials and methods: The AO relocalization in treated-BC cells was recorded using a fluorescence microscopy equipped with a color CCD camerain a real-time fashion. We designed and developed a device equipped with 6 blue-light (443.7 nm) LED corresponding to the center of a 6-well plate. The cell viability was determined using WST-1 reagents, continuous quantification with multi-mode image-based reader, and time-lapse imaging with Cytosmart Systemin human immortalized urothelial (SV-Huc1) and BC cell lines (RT4, 5637 and T24) treated with indicated concentrations of AO or blue-light exposure alone or in combination. Mouse xenograft model bearing T24 cells with or without AO treatment was utilized to investigate the tumor formation in vivo.
Results:TheAO translocation was clearly monitored in fluorescent microscopy with decreased red fluorescent and increased green fluorescent over exposure duration. Treatment ofAO or blue-light exposure alone did not cause a significant decrease of cell viability in BC cells. However, AO exhibited a dose-dependent increment of cytotoxicity toward BC cells with extended duration of blue-light exposure. In addition, this phenomenon was more prominent in human BC cell lines compared to SV-Huc1cells. Furthermore, tumor formation was completely abolished in xenograft model with T24 cells treated with AO and exposure with blue lights. These results suggested that AO, as a photosensitizer, disrupts acidic organelles in BC cells under blue light irradiation.
Conclusion:Blue light irradiation in BC cell treated with AO causes cell death. This finding may serve as a novel therapeutic strategy against human bladder cancer.