黑种草子抑制乳腺癌细胞增殖的组分筛选及机制分析  

Study on the active constituents and anti-proliferation mechanisms of Nigellae Semen

作　　者：王原来[1] 杨晓露[1] 卞卡[2] 章丹丹[1] 

机构地区：[1]上海中医药大学穆拉德中药现代化研究中心,上海201203 [2]美国乔治华盛顿大学生物化学与分子生物学系,华盛顿20052

出　　处：《时珍国医国药》2013年第6期1289-1291,共3页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金青年项目(No.81001666);上海市教育委员会科研创新项目(No.13YZ048);上海高校选拔培养优秀青年教师基金资助项目(No.SZY07029)

摘　　要：目的结合体外筛选确定黑种草子抑制小鼠乳腺癌4T1细胞增殖的有效组分并进行初步的机制分析。方法通过有机溶剂萃取和大孔树脂富集获得黑种草子各提取物,结合体外MTT法筛选黑种草子的各提取物中对4T1细胞具增殖抑制作用的有效组分;集落形成法考察有效组分对细胞集落形成的影响;Western Blot法考察黑种草子有效组分干预前后4T1细胞中p-JAK2的蛋白表达。结果黑种草子总黄酮(相对含量为60.19%)抑制4T1细胞增殖的IC50为最低,同时呈剂量依赖性抑制4T1细胞的集落形成,并下调细胞中p-JAK2的蛋白表达。结论黑种草子总黄酮是黑种草子抑制乳腺癌4T1细胞增殖和集落形成的有效组分,部分是通过抑制JAK/STAT信号转导通路中关键蛋白的磷酸化而发挥抗增殖效果的。Objective To study the active constituents and anti-proliferation mechanism of Nigellae Semen.Methods Ethanol,light petroleum,ethyl acetate,n-butanol and water extraction,different fractions from ethyl acetate and n-butanol were separated by using organic solution and macroporous resin.MTT assay for breast cancer cells proliferation capability.Colony formation assay in three-dimensional culture for cells colony formation calculation.Western blot for protein expression.Results Compared with other constituents,total falvones of Nigellae Semen（TFNS） exhibited the strongest suppressing effect on cell proliferation.TFNS showed dose-dependent inhibitory effect on colony formation.The protein expression of p-JAK2 was inhibited by TFNS,too.Conclusion These results suggest that TFNS may provide potential anti-proliferation and inhibitory effect of colony formation through inhibiting JAK2 protein phosphorylation on JAK/STAT signal pathway activation.

关 键 词：黑种草子 有效组分 乳腺癌 抑制增殖 JAK2 STAT3信号通路 

分 类 号：R285.5[医药卫生—中药学]