为了考察中药制剂西黄丸(Xihuang Pill)对肝癌细胞的作用及对Yes相关蛋白/具有PDZ结合基序的转录共激活因子(Yes-associated protein/transcriptional co-activator with PDZ-binding motif,YAP/TAZ)信号通路的影响,在SD大鼠中利用灌胃给药和动脉采血技术,制备西黄丸含药血清;利用磺酰罗丹明B(sulforho-damine B,SRB)染色技术,检测西黄丸含药血清对肝癌细胞增殖能力和克隆形成能力的影响;利用流式细胞术和原位末端转移酶标记(terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling,TUNEL)技术,检测西黄丸含药血清对肝癌细胞凋亡的影响;利用小室迁移测定实验模型,检测西黄丸含药血清对肝癌细胞侵袭和迁移能力的影响;利用小鼠肿瘤细胞异种移植模型,检测西黄丸对肝癌细胞体内增殖能力的影响;利用蛋白质印迹技术和荧光定量PCR技术,检测西黄丸含药血清对YAP/TAZ信号通路相关蛋白质和基因表达的影响.体外结果显示,西黄丸含药血清显著抑制肝癌细胞的体外增殖、克隆形成、侵袭和迁移能力,显著促进肝癌细胞的凋亡,显著抑制YAP/TAZ信号通路相关蛋白质和基因的表达,且具有剂量依赖性.体内结果显示,西黄丸显著抑制肝癌细胞的体内增殖能力,且具有剂量依赖性.研究结果提示,西黄丸能抑制肝癌细胞的体内外增殖、侵袭和转移能力,并可能在YAP/TAZ信号通路中发挥作用.
Effect and Mechanism of Xihuang Pill on Inhibiting the Proliferation and Invasion of Liver Cancer Cells
In order to investigate the effect of Xihuang Pill on liver cancer cells and Yes-associated protein/transcriptional co-activator with PDZ-binding motif(YAP/TAZ)signaling pathway,the sera containing the pill ingredients were prepared by intragastric administration and arterial blood collection in SD rats.The prolife-ration and clonogenesis of liver cancer cells were detected by sulforhodamine B(SRB)staining.The hepato-ma apoptosis was evaluated using flow cytometry and terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling(TUNEL)method.The invasion and migration abilities of liver cancer cells were tested using the Transwell model,and their in vivo proliferation ability was detected using the xenotransplantation model of mouse tumor cells.The YAP/TAZ signaling pathway related proteins and gene expression were ex-plored by Western-blot and fluorescent quantitative PCR.The in vitro results showed that the sera containing pill ingredients significantly inhibited the proliferation,clonal formation,invasion and migration of liver can-cer cells,significantly promoted the liver cancer apoptosis,and significantly inhibited the expression of YAP/TAZ signaling pathway related proteins and genes in a dose-dependent manner.The in vivo results showed that Xihuang Pill significantly inhibited the proliferation of liver cancer cells in a dose-dependent manner.The study demonstrated that Xihuang Pill can inhibit the in vitro and in vivo proliferation,invasion,and metastasis capabilities of liver cancer cells,and may play a role in the YAP/TAZ signaling pathway.
万方数据
维普
