红毛七总皂苷体外抑制乳腺癌细胞ZR-75-30增殖的研究  

Inhibitory effects of total saponins isolated fromRadix et Rhizoma Leonticis on human breast cancer ZR-75-30 cells

作　　者：康圆 樊梦莹 于润泽 王红英[1] 展颖转[1] KANG Yuan;FAN Mengying;YU Runze;WANG Hongying;ZHAN Yingzhuan(School of Pharmacy,Xi′an Jiaotong University,Xi′an 710061,China)

机构地区：[1]西安交通大学药学院,西安710061

出　　处：《西北药学杂志》2018年第6期752-756,共5页Northwest Pharmaceutical Journal

基　　金：国家自然科学基金资助项目(编号:81302800);陕西省自然科学基础研究计划资助项目(编号:2015JQ8296);陕西省中医管理局中医药科研项目(编号:JCMS028)

摘　　要：目的考察红毛七总皂苷对乳腺癌细胞ZR-75-30增殖、凋亡及迁移的影响及其作用机制。方法采用MTT和平板克隆形成实验,检测红毛七总皂苷对ZR-75-30细胞增殖的影响,通过Hoechst染色与细胞划痕实验,研究红毛七总皂苷对ZR-75-30细胞凋亡及迁移的影响,采用Western Blot检测凋亡及迁移相关蛋白的表达。结果 MTT和平板克隆形成实验结果表明,红毛七总皂苷能显著抑制乳腺癌细胞ZR-75-30的增殖,其在24,48和72h的半数抑制质量浓度(IC50)值依次为0.539 1,0.348 2和0.259 6mg·mL^(-1);Hoechst染色与细胞划痕实验结果表明,红毛七总皂苷能够诱导乳腺癌细胞ZR-75-30凋亡抑制细胞迁移;并进一步通过Western Blot实验证明红毛七总皂苷诱导乳腺癌细胞ZR-75-30凋亡的作用是通过下调抗凋亡蛋白Bcl-2,上调凋亡标志性蛋白Cleaved-Caspase-3和Cleaved PARP的表达实现的,且呈现剂量依赖性;同时证明了红毛七总皂苷能够下调金属基质蛋白酶MMP-2和MMP-9的表达,进而抑制乳腺癌细胞ZR-75-30的迁移。结论红毛七总皂苷能在体外抑制乳腺癌细胞ZR-75-30增殖,通过下调抗凋亡蛋白Bcl-2,上调Cleaved-Caspase-3和Cleaved PARP诱导ZR-75-30细胞凋亡;通过下调MMP-2和MMP-9的表达,抑制ZR-75-30细胞的迁移。Objective The effects of total saponins isolated from Radix et Rhizoma Leonticis （RLTS） on the proliferation,apoptosis and migration of ZR-75-30 cells were investigated. Methods Inhibitory effect of RLTS on ZR-75-30 cells was investigated using MTT assay and cell clony formation assay,while wound scratch assay and Hoechst assay were used to study the effect of RLTS on apoptosis and migration of ZR-75-30 breast cancer cells.The expression of apoptosis and migration associated proteins in ZR-75-30 cells were detected by Western Blot analysis. Results MTT assay and cell colony formation assay demonstrated that RLTS significantly inhibited the growth of ZR-75-30 cells,and the IC 50 values were 0.539 1,0.348 2 and 0.259 6 mg·mL -1 at the time point 24,48 and 72 h respectively.The results of Hoechst assay and wound scratch assay demonstrated that RLTS could induce apoptosis of ZR-75-30 cells in a dose-dependent manner as well as inhibit its migration.Western Blot analysis showed that RLTS could downregulate the expression of Bcl-2,Cleaved-Caspase-3 and Cleaved PARP protein,furthermore,RLTS can inhibit cell migration and downregulate the proteins that mediated metastasis including MMP-2 and MMP-9. Conclusion RLTS could significantly inhibit the growth of ZR-75-30 cells in vitro .Treatment of RLTS could induce apoptosis of ZR-75-30 cells through the downregulation of Bcl-2 and upregulation of Cleaved-Caspase-3 and Cleaved PARP.Meanwhile,the reduced migration of ZR-75-30 cells was due to the downregulation of MMP-2 and MMP-9 by RLTS.

关 键 词：总皂苷 乳腺癌 凋亡 迁移 

分 类 号：R965[医药卫生—药理学]