摘要
目的观察开口箭皂苷对S-180肉瘤细胞荷瘤小鼠NF-κB mRNA的影响,证实开口箭对实体瘤的抑制作用,探讨其分子机制。方法计算四噻唑蓝(MTT)法检测不同浓度的开口箭皂苷对S-180肉瘤细胞体外培养下抑制率,并进行电镜形态观察。同时建立S-180肉瘤荷瘤小鼠模型,给予不同浓度的开口箭皂苷进行干预,以5-氟尿嘧啶(5-Fu)为阳性对照,观察细胞形态,用凝胶电泳迁移率实验(EMSA)技术检测NF-κB的变化;实时荧光定量PCR技术(FQ-PCR)检测各组NF-κB mRNA。结果开口箭皂苷呈浓度依赖性抑制S-180肉瘤细胞增殖,以48h作用最佳。开口箭各组荷瘤裸鼠的瘤质量呈浓度依赖性下降,差异均有统计学意义(均P<0.05),各给药组瘤组织NF-κB p65 mRNA及蛋白表达水平均明显低于空白组(均P<0.05),并随着开口箭皂苷浓度的增加逐渐降低,各组间比较差异均有统计学意义(均P<0.05)。结论开口箭皂苷明显抑制S-180肉瘤细胞体外增值,具有抑制实体肉瘤作用,降低NF-κB mRNA是其分子机制之一。
Objective To study the effect of saponins from Tupistra chinensis Bak （STCB） on the growth of sarcoma S-180 cells in vitro and in mouse xenografts as well as the underlying mechanisms. Methods Cell proliferation was assessed by MTT assay. Cell cycle distribution was determined by flow cytometry. Sarcoma S- 180 tumor-bearing mice were treated with different doses of STCB with 5-fluorouracil （5-Fu） as a positive control. The activity of nuclear factor （NF）-κB was detected by gel mobility shift assay. The mRNA level of NF-κB was determined by real-time quantitative RT-PCR. Results In vitro STCB inhibited the growth of S-180 cells in a concentration-dependent manner, which was accompanied by cell cycle arrest at S-phase. In vivo STCB significantly inhibited the growth of S-180 tumor mouse xenografts in a dose-dependent manner with apparent induction of cell apoptosis. Moreover, STCB inhibited the activity of NF-κB p65 and reduced the expression of NF-κB p65 mRNA in mouse xenografts. Conclusions STCB inhibits the proliferation and cell cycle progression of S- 180 cells by suppressing NF-κB signaling in mouse xenografts. Our findings suggest STCB is a promising agent for the treatment of sarcoma.
作者
叶太生
张莹雯
王秀萍
张贤梅
张曼玲
YE Taisheng;ZHANG Yingwen;WANG Xiuping;ZHANG Xianmei;ZHANG Manling(Department of Integrated Traditional Chinese and Westem Medicine, Zhongnan Hospital of Wuhan University, Wuhan 430071;Department of Integrated Traditional Chinese and Western Medicine, The Third Hospital ofWuhan, Wuhan 430060)
出处
《湖北中医药大学学报》
2018年第2期5-9,共5页
Journal of Hubei University of Chinese Medicine
基金
湖北省自然科学基金(项目编号:2009CHB001)
