喹唑啉酮衍生物MJ-56經由光毒性抑制膀胱癌細胞
陳宏恩1、林致凡4、林宜佳1,3、蔡德甫1,3、仇光宇1,3、黃一勝1,2,3
1新光醫院 外科部 泌尿科; 2台北醫學大學醫學系 泌尿學科; 3輔仁大學醫學系 泌尿學科;
4新光醫院中央實驗室
A novel quinazolinone derivative, MJ-56, exhibits phototoxicity toward human bladder cancer cells
Hung-En Chen1#, Ji-Fan Lin4#,Yi-Chia Lin1,3,Te-Fu Tsai1,3, and Kuang-Yu Chou1,3, Thomas I-Sheng Hwang1,2,3
1Department of Urology, Shin Kong Wu Ho-Su Memorial Hospital, Department of Urology,
2Taipei Medical University, Division of Urology, 3School of Medicine, Fu-Jen Catholic University, 4Central Laboratory, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan.
# these authors contributed equally to this work
Abstract
Background: Quinazolinones 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 the novel quinazolinone 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 bladder cancer 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 in both 5637 and T24 cells. However, treatment of 0.125 μM MJ-56 for 1-hour and exposed to blue light for 30 seconds significantly reduced cell viability in 5637 and T24 cells. 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 compared to bladder cancer 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 bladder cancer cells.
Conclusions: 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.