文章摘要
郭檬檬,应语,陈海东,等.山茱萸多糖通过AMPK/SIRT1通路抑制乳腺癌生长作用机制研究[J].江苏中医药,,():.
山茱萸多糖通过AMPK/SIRT1通路抑制乳腺癌生长作用机制研究
Study on the Mechanism of Cornus Polysaccharides Inhibiting Breast Cancer Growth Through AMPK/SIRT1 Pathway
投稿时间：2023-08-10  修订日期：2024-04-03
DOI：
中文关键词: 山茱萸多糖  腺苷酸活化蛋白激酶/沉默信息调节因子1通路  乳腺癌细胞  增殖  迁移
英文关键词: cornus polysaccharide  adenylate-activated protein kinase/silent information regulator 1 pathway  breast cancer cells  proliferation  Migration
基金项目:国家自然科学基金(编号:81873305,负责人:姚昶)
作者单位邮编
郭檬檬 南通市中医院 226000
应语 江苏省中医院 220000
陈海东 南通市中医院 226000
缪翠维 南通市中医院 226000
姚昶* 江苏省中医院 220000
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中文摘要:
      目的：探讨山茱萸多糖对乳腺癌细胞增殖、迁移的影响及作用机制。方法：体外培养MCF-7细胞并分为对照组、山茱萸多糖低、中、高剂量组（浓度分别为12.5、25.0、50.0 mg/mL）和山茱萸多糖高剂量+腺苷酸活化蛋白激酶（AMPK）激活剂（AICAR）组（50.0 mg/mL山茱萸多糖+2.0 mmol/L AICAR），平板克隆形成实验检测MCF-7细胞增殖，流式细胞术检测细胞凋亡，划痕实验检测细胞迁移，Transwell法检测细胞侵袭，Western blot实验检测AMPK、磷酸化AMPK（p-AMPK）、沉默信息调节因子1（SIRT1）蛋白表达。建立荷瘤小鼠模型检验山茱萸多糖对乳腺癌移植瘤生长的影响。结果：山茱萸多糖低、中、高剂量组的MCF-7细胞集落形成数低于对照组（P＜0.05），山茱萸多糖高剂量+AICAR组MCF-7细胞集落形成数高于山茱萸多糖高剂量组（P＜0.05）。山茱萸多糖低、中、高剂量组的MCF-7细胞凋亡率高于对照组（P＜0.05），山茱萸多糖高剂量+AICAR组MCF-7细胞凋亡率低于山茱萸多糖高剂量组（P＜0.05）。山茱萸多糖低、中、高剂量组的MCF-7细胞凋亡率高于对照组（P＜0.05），山茱萸多糖高剂量+AICAR组MCF-7细胞凋亡率低于山茱萸多糖高剂量组（P＜0.05）。山茱萸多糖低、中、高剂量组的MCF-7细胞划痕愈合率、细胞侵袭数低于对照组（P＜0.05），山茱萸多糖高剂量+AICAR组MCF-7细胞划痕愈合率、细胞侵袭数高于山茱萸多糖高剂量组（P＜0.05）。山茱萸多糖低、中、高剂量组的MCF-7细胞p-AMPK、SIRT1蛋白表达低于对照组（P＜0.05），山茱萸多糖高剂量+AICAR组MCF-7细胞p-AMPK、SIRT1蛋白表达高于山茱萸多糖高剂量组（P＜0.05）。山茱萸多糖各剂量组对上述各指标的作用呈剂量依赖性。山茱萸多糖组移植瘤体积、质量与瘤体组织p-AMPK、SIRT1蛋白阳性表达率低于对照组（P＜0.05）。结论：山茱萸多糖可能通过抑制AMPK/SIRT1通路激活抑制乳腺癌肿瘤生长。
英文摘要:
      Objective: To investigate the impacts and mechanism of Cornus polysaccharide on the proliferation and migration of breast cancer cells. Methods: MCF-7 cells cultured in vitro were grouped into control group, Cornus polysaccharide low, medium and high dose groups (concentrations of 12.5, 25.0, 50.0 mg/mL, respectively), and Cornus polysaccharide high dose + adenylate-activated protein kinase (AMPK) activator (AICAR) group (50.0 mg/mL Cornus polysaccharide + 2.0 mmol/L AICAR), MCF-7 cell proliferation was detected by plate clone formation assay, apoptosis was detected by flow cytometry, cell migration was detected by scratch assay, cell invasion was detected by Transwell assay, and protein expressions of AMPK, phosphorylated AMPK (p-AMPK) and silent information regulator 1 (SIRT1) were detected by Western blot assay. tumor bearing mouse model was established to test the effect of cornus polysaccharide on the growth of breast cancer transplanted tumor. Results: The colony formation number of MCF-7 cells in Cornus polysaccharide low, medium and high dose groups was lower than that in control group (P<0.05), and the colony formation number of MCF-7 cells in Cornus polysaccharide high dose +AICAR group was higher than that in Cornus polysaccharide high dose group (P<0.05). The apoptosis rate of MCF-7 cells in Cornus polysaccharide low, medium and high dose groups was higher than that in control group (P<0.05), and the apoptosis rate of MCF-7 cells in Cornus polysaccharide high dose +AICAR group was lower than that in Cornus polysaccharide high dose group (P<0.05). The apoptosis rate of MCF-7 cells in Cornus polysaccharide low, medium and high dose groups was higher than that in control group (P<0.05), and the apoptosis rate of MCF-7 cells in Cornus polysaccharide high dose +AICAR group was lower than that in Cornus polysaccharide high dose group (P<0.05). The scratch healing rate and cell invasion number of MCF-7 cells in Cornus polysaccharide low, medium and high dose groups were lower than those in control group (P<0.05), and the scratch healing rate and cell invasion number of MCF-7 cells in Cornus polysaccharide high dose +AICAR group were higher than those in Cornus polysaccharide high dose group (P<0.05). The protein expressions of P-AMPK and SIRT1 in MCF-7 cells in Cornus polysaccharide low, medium and high dose groups were lower than those in control group (P<0.05), and the protein expressions of P-AMPK and SIRT1 in MCF-7 cells in Cornus polysaccharide high dose +AICAR group were higher than those in Cornus polysaccharide high dose group (P<0.05). The effects of polysaccharides of Cornus officinalis in each dose group on the above indexes were dose-dependent. The graft volume, weight and the positive expression rates of P-AMPK and SIRT1 protein in the tumor tissue of Cornus polysaccharide group were lower than those of control group (P<0.05). Conclusion: Cornus polysaccharide inhibits growth of breast cancer by inhibiting the activation of AMPK/SIRT1 pathway.
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