Li Jin-Quan, Chen Rui-Chuan, Cai Ke-Xia, Ye Zhi-Yun
Department of Biology, Life Science School of Xiamen University, Xiamen, Fujian, 361005, P.R.China.
Ai Zheng. 2003 Mar;22(3):253-6.
BACKGROUND & OBJECTIVE: Photochemical riboflavin, the production of reactive oxygen species (ROS), has been reported having cytotoxity on some cancer cells. The aim of this study was to investigate the effect of photochemical riboflavin on inducing apoptosis in human gastric cancer cell line MGC80-3.
Trypan blue exclusion method, Giemsa staining, DNA electrophoresis, DNA quantification, Western blot analysis, and flow cytometry were conducted to determine the effect of photochemical riboflavin on cell survival, morphology, DNA fragmentation, gene expression, and cell cycle arresting.
The cell viability dropped down according to the riboflavin concentration and treating time. Exposure of the cells in 20 micromol/L riboflavin for 24 hours resulted in typical apoptotic morphology and G(2)/M arresting. As MGC80-3 cells were separately treated with 10, 20, 30, and 40 micromol/L photochemical riboflavin, DNA electrophoresis showed that the ladder bands, a typical feature of apoptotic cell, appeared in the groups treated by over 20 micromol/L photochemical riboflavin. The efficiency rates of DNA fragmentation were 35.4%, 54.1%, 70.6%, and 86.8%, respectively. Western blot analysis showed that the expression of apoptosis related proteins p53, C-myc, and Bax were up-regulated whereas the expression of Bcl-2 was down-regulated.
These results indicate that photochemical riboflavin has high efficiency of inducing cell apoptosis on MGC 80-3 cells in vitro and there is a correlation between apoptosis and G(2)/M arresting.
光化学核黄素可产生活性氧(ROS),已有报道称其对某些癌细胞具有细胞毒性。本研究旨在探讨光化学核黄素对人胃癌细胞系MGC80 - 3诱导凋亡的作用。
采用台盼蓝排斥法、吉姆萨染色、DNA电泳、DNA定量、蛋白质免疫印迹分析及流式细胞术,以确定光化学核黄素对细胞存活、形态、DNA片段化、基因表达及细胞周期阻滞的影响。
细胞活力随核黄素浓度和处理时间下降。将细胞暴露于20 μmol/L核黄素中24小时,导致典型的凋亡形态及G(2)/M期阻滞。当MGC80 - 3细胞分别用10、20、30和40 μmol/L光化学核黄素处理时,DNA电泳显示,在光化学核黄素浓度超过20 μmol/L处理的组中出现了凋亡细胞的典型特征——梯状条带。DNA片段化的效率分别为35.4%、54.1%、70.6%和86.8%。蛋白质免疫印迹分析显示,凋亡相关蛋白p53、C - myc和Bax的表达上调,而Bcl - 2的表达下调。
这些结果表明,光化学核黄素在体外对MGC 80 - 3细胞具有高效诱导细胞凋亡的作用,且凋亡与G(2)/M期阻滞之间存在相关性。