摘要
目的探讨连翘抗肿瘤活性成分LQ-4体外对人宫颈癌HeLa细胞的增殖抑制和诱导HeLa细胞凋亡及机制。方法通过水提取、醇沉、大孔吸附树脂柱层析分离、薄层层析,分离提取LQ-4;四甲基偶氮唑蓝比色法观察LQ-4对HeLa细胞增殖抑制作用;吖啶橙/溴化乙锭双荧光染色法、透射电镜观察细胞凋亡的形态学变化;west-ern blot法测定HeLa细胞caspase-8表达水平。结果 LQ-4体外对HeLa细胞的生长、增殖均有抑制作用,12、24和48 h的半数抑制浓度(IC50)分别为93.74、33.30、22.65μg/mL;LQ-4作用HeLa细胞后,透射电镜观察可见典型凋亡细胞形态;LQ-4作用于HeLa细胞后,促进caspase-8酶原活化,在43及18 kD上均有表达。结论 LQ-4对HeLa细胞的增殖具有抑制作用,可诱导HeLa细胞凋亡,其机制可能与caspase-8蛋白酶原裂解有关。
Objective To study the cell inhibitory and apoptosis effect of bioactive compounds extracted from For- sythia suspensa （LQ-4） on HeLa cells in vitro, and to study the mechanism of anti-tumor activity of LQ-4. Methods LQ-4 was extracted by means of alcohol precipitation, macroporous resin chromatography, and thin-layer chromatogra- phy. Methyl thiazolyl tetrazolium（MTT） assay was used to evaluate the growth inhibition rate of HeLa cells treated by different concentrations of LQ-4. Arcridine orange/ethidium bromide （AO/EB） fluorescent staining and transmission electron microscopy（TEM） were used to observe cell morphology. Western blot was used to detect the cell apoptosis-re- lated protein in HeLa cells treated by LQ-4 （50 g/ml） at different times. Results The anti-tumor active components （LQ-4） showed time-dose-dependent inhibitory effect on the proliferation of HeLa cells in vitro. The concentration for 50% inhibition（ IC50 ） at 12,24, and 48 hours was 93.74,33.30, and 22. 65 μg/mL, respectively. Fluorescence micro- scope and TEM detection showed that HeLa cells presented characteristic morphological changes of apoptosis after LQ-4 treatment. The results of western blot revealed that LQ-4 （50 μg/mL） could promote the activation of the zymogen of caspase-8. Conclusion LQ-4 has an inhibitory effect on HeLa cell proliferation and could induce HeLa cell apoptosis, which is probably related to the decomposition of caspase-8 protease.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2013年第3期397-399,共3页
Chinese Journal of Public Health
