新型喹唑啉衍生物MJ-56為綠螢光敏染劑並在人類膀胱癌細胞中誘發光毒性
陳宏恩1#、林致凡2#、蔡德甫1、楊尚哲2、林宜佳1,4、仇光宇1、黃一勝1,2,3,4
1新光醫院 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
MJ-56, a novel quinazolin derivative, emits green fluorescent and exhibits phtotoxicity toward human bladder cancer cells
Hung-En Chen1#, Ji-Fan Lin2#, Te-Fu Tsai 1, Shan-Che Yang2, Yi-Chia Lin1,4, Kuang-Yu Chou1, and Thomas I-Sheng Hwang1,2,3,4
Department of Urology, Shin Kong Wu Ho-Su Memorial Hospital1, Central Laboratory, Shin Kong Wu Ho-Su Memorial Hospital2, Department of Urology, Taipei Medical University3, Division of Urology, School of Medicine, Fu-Jen Catholic University4, Taipei, Taiwan.
# These authors contributed equally to this work.
Purpose: Quinazolines which processing a wide spectrum of biological properties such as antibacterial, antifungal, antivirus, and anticancer activities, are considered as one of the most important heterocycles in medicinal chemistry. The derivatives of quinazolinone are known to inhibit cell metastasis and promote cell death in various types of cancer. Here, we described for the first time that a novel quinazoline derivative MJ-56 (6-pyrrolidinyl-2-(3-bromostyryl)quinazoline-4-one) emitted green fluorescent in the cytosol and exhibits phototoxicity toward human bladder cancer cells (5637 and T24) upon blue-light exposure.
Materials and Methods: Human BC cell lines (5637 and T24) and immortalized uroepithelial cell line (SV-Huc-1) were used in this study. Observation of the fluorescent in MJ-56 treated cells with or without MitoTracker or LysoTracker was conducted by using a fluorescent microscopy. Cells were treated with indicated concentrations of MJ-56 for 1 hours with or without the blue light irradiation. The cell viability was detected by (1) WST-1 reagent immediately 1-hour post-treatment, (2) Cytosmart System that records the cell morphology and calculates cell viability automatically for 36 hours, and (3) colony formation assays. The ADT/ATP levels in MJ-56 treated cells were detected using a commercial kit.
Results: We found that green fluorescent was readily detectable in the cytosol of cells treated with 0.125 μM MJ-56 for only 1-hour. Vital staining of mitochondria or lysosomes using MitoTracker or LysoTracker, respectively, demonstrated that the fluorescent caused by MJ-56 was not located in either organelles. Administration of MJ-56 for 24 hours did not cause significant loss of cell viability, however, 0.125 μM MJ-56 treated for 1-hour and exposed to blue-light for 30 seconds significantly reduced cell viability. To understand the impact of MJ-56 on normal urothelial cells, we performed the same treatment using SV-Huc-1, an immortalized urothelial cell line. The results showed that MJ-56 has small effect on SV-Huc-1 cells, even with the blue-light exposure. The caspase 3/7 activities in cells treated with MJ-56 and exposed to blue-light were significantly increased at 1-hour post treatment. However, the DNA fragmentation cannot be detected at 1, 6, or 24 hours-post-treatment due to the significantly loss of viable cells. The increased ADP/ATP ratio in treated cells suggested that MJ-56 may induce necrosis rather than apoptosis in BC cells.
Conclusion: W Our results demonstrated that MJ-56 exhibits phototoxicity to bladder cancer cells with minimal impact on urothelial cells, indicating a potential novel therapeutic strategy against bladder cancer. However, the mechanism underlying MJ-56 induced cell death as well as the translational studies warrant further investigation.