薯蓣皂苷元对人宫颈癌细胞增殖的影响(1)
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[摘要] 目的 研究薯蓣皂苷元对人宫颈癌细胞增殖的影响。方法 培养人宫颈癌细胞株Hela，不同浓度的薯蓣皂苷元干预24、48、72h，采用MTT、EdU检测细胞增殖，流式细胞术检测细胞周期，Western Blot检测细胞周期蛋白Cyclin D1、Cyclin B1。结果 薯蓣皂苷元在20μmol/L浓度时抑制Hela细胞活力(P<0.01)，呈现量效和时效特征；薯蓣皂苷元作用后EdU阳性细胞明显减少，薯蓣皂苷元引起G0/G1期细胞阻滞，减少S期细胞，降低Cyclin D1蛋白表达，而对Cyclin B1蛋白表达没有明显影响。结论 薯蓣皂苷元通过阻滞细胞G0/G1期来抑制人宫颈癌Hela细胞株的增殖。
[关键词] 薯蓣皂苷元；宫颈癌；细胞增殖；细胞周期
[中图分类号] R737.33 [文献标识码] A [文章编号] 1673-9701(2011)26-05-03
Influence of Diosgenin on Cell Proliferation in Human Cervical Cancer
ZHANG Wenxiu
Department of Obstetrics and Gynecology， the First People's Hospital of Jining City， Jining 272100， China
[Abstract] Objective To evaluate the possible effects of diosgenin on cell proliferation in human cervical cancer. Methods The Hela cells were treated with different concentrations of diosgenin for 24， 48 and 72 hours， respectively. Cell proliferation was tested by MTT and EdU assay， and cell cycle analysis was performed by FACS. In addition， Cyclin D1 and B1 protein expressions were tested by Western Blotting， respectively. Results We found that 20 μmol/L diosgenin began to markedly inhibit cell viability (P<0.01) and there were dose-dependent and time-dependent effects of diosgenin on cell viability. We found that the number of EdU+ cells (EdU-labeled replicating cells) was significantly reduced in diosgenin treatment group. Diosgenin decreased expression of cyclin D1 and arrested them at the G0/G1 phase. Conclusion Diosgenin inhibits the proliferation of human cervical cancer cells by blocking them at the G0/G1 phase.
[Key words] Diosgenin； Cervical cancer； Cell proliferation； Cell cycle
现代药理研究表明薯蓣皂苷及其衍生物具有免疫调节、降血脂、抗衰老、抗氧化、抗关节炎、保护胃黏膜、祛痰、溶血、脱敏、灭钉螺、抗肿瘤、抗艾滋病、抗血小板聚集、增强心脏收缩力、减慢心率、抗动脉硬化、改善微循环等多种药理活性[1，2]。宫颈癌是妇科最常见的恶性肿瘤，发病率呈逐年上升和年轻化趋势；近年来，国内外对化疗在宫颈癌的应用进行基础和临床方面研究，取得较大进展。本研究观察薯蓣皂苷元对人宫颈癌Hela细胞株增殖和细胞周期的影响，探讨其对治疗宫颈癌的可能作用。
1 材料与方法
1.1 细胞培养
人宫颈癌细胞株购自上海瑞齐生物科技有限公司，用含10%新生牛血清(NBS，Gibco，USA)、100U/mL青霉素和100μg/mL链霉素、2mmol/L谷氨酰胺的DMEM培养基(Gibco，USA)培养；细胞培养孵箱内培养条件：5%CO2，95%空气，温度37℃，湿度98%。细胞在培养皿中密度达到70%～80%时，进入实验干预。
1.2 薯蓣皂苷元贮存液配制
原粉为美国Sigma公司产品，按照说明书要求，溶于酒精配制100mM贮存液，无菌分装-80℃保存备用。工作液酒精浓度0.5%。分组实验，无菌吸取贮存液加入饥饿培养液中 ......
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