Lin Min, Xie Hai-Long, Su Qi, Zhou Jian-Guo, Tan Hui, Yi Lan, Tang Hai-Lin, Huang Yan, Shi Ying
Cancer Research Institute, Nanhua University, Hengyang, Hunan, 421001, PR China.
Ai Zheng. 2007 Apr;26(4):351-6.
BACKGROUND & OBJECTIVE: Diallyl disulfide (DADS) can induce apoptosis in various cancer cell lines in vitro. Leukemia is the most common pediatric malignancy. Recent studies have suggested that DADS can induce apoptosis in human leukemia cells, but its mechanisms remain unclear. This study was to investigate the biological effects of DADS on the apoptosis of human leukemia cell line HL-60, and explore its molecular mechanisms.
After treatment of DADS, cell apoptosis was verified by flow cytometry with Annexin V/PI staining, DNA agarose gel electrophoresis, transmission electron microscopy. The expression profile of apoptosis-associated genes in HL-60 cells, with or without 4-hour treatment of DADS (60 micromol/L), was identified by gene array. The up-regulated Fas-L gene and down-regulated Bag-1 gene were confirmed by reverse transcription-polymerase chain reaction (RT-PCR).
When treated with 15, 30, 60, 120 micromol/L DADS for 24 h, HL-60 cells presented obvious subdiploid peaks. When treated with 60 micromol/L DADS for 4, 8, 12, 24 h, early apoptotic cells were greatly increased. When treated with 60 micromol/L DADS for 24 h, DNA extracted from HL-60 cells displayed a characteristic ladder pattern on agarose gel electrophoresis; typical morphologic apoptotic changes were observed under electron microscope, including cell shrinkage, nuclear condensation, and formation of apoptotic bodies; the differential expression of 8 apoptosis-associated genes were found with gene array. The expression of Fas-L and Bag-1 detected by RT-PCR were consistent with those detected by gene array.
DADS could induce apoptosis in HL-60 cells, which might be mediated by some specific genes and various signal transduction pathways.
二烯丙基二硫化物（DADS）在体外可诱导多种癌细胞系发生凋亡。白血病是儿童最常见的恶性肿瘤。近期研究表明，DADS可诱导人白血病细胞凋亡，但其机制尚不清楚。本研究旨在探讨DADS对人白血病细胞系HL-60凋亡的生物学效应，并探索其分子机制。
用DADS处理后，通过Annexin V/PI染色流式细胞术、DNA琼脂糖凝胶电泳、透射电子显微镜检测细胞凋亡。采用基因芯片技术鉴定HL-60细胞在有无4小时DADS（60微摩尔/升）处理情况下凋亡相关基因的表达谱。通过逆转录聚合酶链反应（RT-PCR）验证Fas-L基因上调和Bag-1基因下调。
用15、30、60、120微摩尔/升DADS处理24小时后，HL-60细胞出现明显的亚二倍体峰。用60微摩尔/升DADS处理4、8、12、24小时后，早期凋亡细胞显著增加。用60微摩尔/升DADS处理24小时后，从HL-60细胞提取的DNA在琼脂糖凝胶电泳上呈现特征性梯状条带；在电子显微镜下观察到典型的凋亡形态学改变，包括细胞皱缩、核浓缩和凋亡小体形成；基因芯片检测发现8个凋亡相关基因表达存在差异。RT-PCR检测的Fas-L和Bag-1表达与基因芯片检测结果一致。
DADS可诱导HL-60细胞凋亡，其机制可能由某些特定基因和多种信号转导通路介导。
