BACKGROUND: Uterine cervix cancer is one of the diseases to damage female health and degrade quality of life. It has been a hot spot to decrease the side effects of radiotherapy and chemotherapy and anti-tumor medicine by using Chinese medicine in medical domain. In Europe, silymarin had been used to treat hepatitis. It had little toxic action and large contents in Chinese herb. Therefore, many researchers are studying its anti-tumor effects now.OBJECTIVE: To study the pharmacological mechanism of silymarin on human cervical cancer cell.DESIGN: The cells cultured in 10% serum medium served as negative control group in vitro.MATERIALS: The experiment was finished in the China-Japan Research Institute of Medical and Pharmaceutical Sciences in Shenyang Pharmaceutical University; Department of Pathological Science of Showa Medical UniversityMATERIASLS: Human cervical cancer cell (HeLa) is the preservative cell line of American ATCC in the Lab. The experiment was completed in the Cell Lab of China-Japan Research Institute of Medical and Pharmaceutical Sciences in Shenyang Pharmaceutical University from Feburary 2003 to July 2003.METHODS: HeLa cell death ratio was measured by methyl thiazol tetrazolium(MTT) assay. Cellular morphologic changes were observed by phase-contrast and fluorescence microscopy. Cell cycle distribution was analyzed by flow cytometry.MAIN OUTCOME MEASURES: The changes of cell death ratio and cell cycle distribution.RESULTS: Silymarin-induced HeLa cell death was relevant to the content of serum in medium. At the does of 80 μmol /L silymarin, the cell death ratio in serum-free group reached 85.27%. The cell death ratio in 5% and 10%serum groups was 35.53% and 7.71% respectively. Morphological changes of HeLa cells demonstrated that silymarin induced HeLa cell death through apoptotic pathway at lower concentration in serum-free condition. Flow cytometry analysis showed that silymarin induced 70. 04% cells death, most of which were in G0/G1 phase.CONCLUSION: Silymarin mainly induce G0/G1 phase HeLa cell death through apoptotic pathway in serum-free medium.%背景:宫颈癌是危害女性身体健康,降低生活质量的疾病之一.利用中药治疗宫颈癌,降低放射治疗、应用抗肿瘤药物治疗副作用对机体伤害已成为医药界的研究热点.在欧洲,水飞蓟素曾广泛用于临床的肝炎治疗.它毒性小,在植物中含量高.因此,一些学者正在研究其抗肿瘤的活性.目的:探讨水飞蓟素对人子宫颈癌细胞的药理作用机制.设计:以体外10%血清正常培养的细胞作为阴性对照的实验对照研究.单位:沈阳药科大学中日医药研究所,昭和药科大学病态科学教研室.材料:人的宫颈癌HeLa细胞为本实验室保存的美国ATCC细胞系.实验于2003-02/07在沈阳药科大学中日医药研究所细胞室完成.方法:噻唑蓝还原法(MTT)分析HeLa细胞的死亡率.采用相差和荧光显微镜观察细胞形态学改变.用流式细胞术分析了细胞周期的变化.主要观察指标:细胞死亡率和细胞周期分化的变化.结果:水飞蓟素诱导的HeLa细胞死亡与培养液中血清含量有关.给予80 μmol/L的水飞蓟素作用,去血清组的细胞死亡率达到85.27%,含5%,10%血清组的死亡率为35.53%,7.71%.细胞的形态学变化表明水飞蓟素诱导的HeLa细胞发生凋亡.流式细胞分析结果显示:水飞蓟素诱导70.04%的细胞发生凋亡,这些凋亡细胞主要来自G0/G1期.结论:水飞蓟素在去血清条件下,诱导处于G0/G1期的HeLa细胞发生凋亡.
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