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

关键词:  西黄丸  沉默调节蛋白 1/性别决定区Y框蛋白 2 信号通路  乳腺癌  增殖  迁移

Key words:  Xihuang Pill  silence regulatory protein 1/sex determining region Y box protein 2 signal pathway  breast cancer  proliferation  transfer

摘要:

[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（6 μmol/L）、西黄丸高剂量+SRT 1720 组（15 g/L+6 μmol/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。

Abstract:

[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 1 720（SIRT1 activator） group （6 μmol/Ｌ），high dose Xihuang pill+SRT 1720 group （15 g/L+6 μmol/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 1 720 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.