摘要
目的:探讨SW体外诱导人胃癌细胞株SGC7901凋亡的机制。方法:应用MTT法确定SW对体外培养的SGC7901细胞的作用剂量,通过流式细胞术及凋亡相关调控基因p53、cmyc、Bcl2的检测对细胞凋亡及细胞周期的测定,观察SW对胃癌细胞SGC7901增殖周期的影响以及抑制肿瘤细胞增殖的方式,同时利用激光共聚焦显微镜对细胞内Ca2+浓度的监测,研究SW与细胞内Ca2+超载的关系。结果:SW体外抗SGC7901细胞的完全致死剂量为6.2μg·ml-l,高于0.05μg·ml-l时抑制作用明显(P<0.05),其LC50为0.84μg·ml-l;经SW0.5～1.5μg·ml-l处理24h可引起凋亡抑制基因p53、Bcl2的明显下降、凋亡促进基因cmyc的明显升高以及肿瘤细胞内Ca2+超载,最终诱导SGC7901细胞凋亡。实验还证实SW主要作用于肿瘤细胞的S期,使瘤细胞主要积聚在S期。结论:SW通过多种途径诱导细胞凋亡可能是其发挥抗癌作用的重要机制。
AIM： To investigate the effects and mechanisms of swainsonine-indueed apoptosis on SGC-7901 cells. METHODES： After being treated with swainsonine, effective dose and median inhibition concentration （IC50） of swainsonine to SGC-7901 cells were examined by MTr assay. Cell cycle distribution and apoptotic rates were analyzed by flow cytometry. Expression of p53, c-myc and Bcl-2 were determined by immunocytochemical method, and the concentration of Ca^2＋ intra-cellular （ [Ca^2＋]i ） was measured by the laser scanning confocal microscope （LSCM）. RESULTS： Swainsonine inhibited cell growth of SGC-7901 in vitro, IC50 of 24h was 0.84μg·ml^-1, and complete inhibition concentration of swainsonine was 6.2μg·ml^-1. Treated with swainsonine at the concentrations of 0.5, 1.5 and 4.5μg·ml^-1 for 24h, the expression of apoptosis inhibiting gene p53 and bcl-2 decreased, and apoptotic trigger gene c-myc increased （ P〈0.05）, as well as [ Ca^2＋ ]i overloading, SGC-7901 cell was induced to apoptosis in the end. The percentage of S phase were 38.8%, 39.7% and 29.6%, respectively （20.0% in control group and 23.2% in 5-Fu group）, the percentage of G2/M phase were 4.5%, 1.7% and 5.3%, respectively （5.5% in control group and 9.0% in 5-Fu group）, and the percentage of G1/M phase was not altered. SGC-7901 cells were treated by swainsonine at the concentrations of 0.5, 1.5 and 4.5μg·ml^-1 for 24h. Compared with the control group, the percentage of S phase were increased and that of G2/M cells were decreased significantly in treatment groups （P〈0.01）. CONCLUSION： Swainsonine can inhibit the cell proliferation and induce apoptosis of SGC-7901 cells, the mechanisms of swainsonine-induced apoptosis may related with [Ca^2＋];overloading and expression of apoptosis-related genes.
出处
《中国临床药理学与治疗学》
CAS
CSCD
2005年第9期978-983,共6页
Chinese Journal of Clinical Pharmacology and Therapeutics
关键词
苦马豆素
胃癌
凋亡
流式细胞仪
细胞周期
激光共聚焦显微镜
swainsonine
gastric carcinoma
apoptosis
flow cytometry
cell cycle
laser scanning confocal microscope
[Ca^2＋]i
