基于SIRT1/SOX2信号通路探究西黄丸抗乳腺癌作用及机制

Study on the anti breast cancer effect and mechanism of Xihuang Pill based on SIRT1/SOX2 signal pathway

[目的]基于沉默调节蛋白1(SIRT1)/性别决定区Y框蛋白2(SOX2)信号通路探究西黄丸抗乳腺癌作用及机制。[方法]将MDA-MB-231细胞分为对照组、西黄丸低剂量组(5 g/L)、西黄丸中剂量组(7.5 g/L)、西黄丸高剂量组(15 g/L)、SIRT1激活剂(SRT 1720)组(6μmol/L)、西黄丸高剂量+SRT 1720组(15 g/L+6μmol/L);CCK-8法和平板克隆实验检测细胞增殖;划痕实验检测细胞迁移;Transwell法检测细胞侵袭;蛋白免疫印迹法(Western Blot)检测细胞中增殖细胞核抗原(PCNA)、基质金属蛋白酶(MMP)-2、MMP-9、SIRT1、SOX2蛋白表达;体内移植瘤实验检测各组裸鼠肿瘤体积、质量及PCNA、MMP-2、MMP-9、SIRT1、SOX2蛋白表达。[结果]西黄丸可抑制MDA-MB-231细胞增殖、迁移、侵袭及裸鼠体内移植瘤的生长,且呈剂量依赖性;SIRT1激活剂SRT 1720促进MDA-MB-231细胞增殖、迁移、侵袭及裸鼠体内移植瘤的生长;高剂量西黄丸加SIRT1激活剂组可以减弱高剂量西黄丸对MDA-MB-231细胞增殖、迁移、侵袭及裸鼠体内移植瘤的生长的抑制作用;同时西黄丸可呈剂量依赖性地抑制MDA-MB-231细胞及裸鼠移植瘤中SIRT1、SOX2蛋白表达,抑制SIRT1/SOX2信号通路活性,并通过降低PCNA、MMP-2、MMP-9表达来抑制MDA-MB-231细胞增殖、迁移、侵袭及裸鼠体内移植瘤的生长。[结论]西黄丸可能通过抑制SIRT1/SOX2信号通路抑制MDA-MB-231细胞增殖、迁移、侵袭及裸鼠体内移植瘤生长。

[Objective]To explore the anti-breast cancer effect and mechanism of Xihuang pill based on silencing regulatory protein 1(SIRT1)/sex determination region Y frame protein 2(SOX2)signaling pathway.[Methods]MDA-MB-231 cells were grouped into control group,low dose Xihuang pill group(5 g/L),medium dose Xihuang Pill group(7.5 g/L),high dose Xihuang Pill group(15 g/L),SRT 1720(SIRT1 activator)group(6μmol/L),high dose Xihuang pill+SRT 1720 group(15 g/L+6μmol/L);CCK-8 method and plate cloning test were used to detect cell proliferation;cell migration was detected by scratch test;transwell method was used to detect cell invasion;Western blot was used to detect the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase(MMP-2),MMP-9,SIRT1,and SOX2 in cells;the tumor volume,mass,quality and PCNA,MMP-2,MMP-9,SIRT1,SOX2 protein expression of nude mice in each group were measured in vivo tumor transplantation experiment.[Results]Xihuang Pill inhibited the proliferation,migration and invasion of MDA-MB-231 cells and the growth of transplanted tumor in nude mice in a dose-dependent manner;SIRT1 activator SRT 1720 promoted the proliferation,migration and invasion of MDA-MB-231 cells and the growth of transplanted tumor in nude mice;the high dose Xihuang Pill plus SIRT1 activator group attenuated the inhibitory effect of high dose Xihuang Pill on the proliferation,migration,invasion of MDA-MB-231 cells and the growth of transplanted tumor in nude mice.At the same time,Xihuang Pill inhibited the expression of SIRT1 and SOX2 proteins in MDA-MB-231 cells and transplanted tumor in nude mice in a dose-dependent manner,inhibited the activity of SIRT1/SOX2 signal pathway,and inhibited the proliferation,migration,invasion of MDA-MB-231 cells and the growth of transplanted tumor in nude mice by reducing the expression of PCNA,MMP-2 and MMP-9.[Conclusion]Xihuang Pill may inhibit the proliferation,migration and invasion of MDA-MB-231 cells and the growth of transplanted tumor in nude mice by inhibiting SIRT1/SOX2 signal pathway.