小檗碱抑制胃癌细胞增殖、迁移及侵袭的作用机制研究  

Mechanism of berberine inhibiting the proliferation,migration and invasion of gastric cancer cells

作　　者：肖小乔 菅佳宁 尚艺婉 周哲旭 陈银芝 李晨 王心仪 陈玉龙[1,2] XIAO Xiao-qiao;JIAN Jia-ning;SHANG Yi-wan;ZHOU Zhe-xu;CHEN Yin-zhi;LI Chen;WANG Xin-yi;CHEN Yu-long(Henan University of Traditional Chinese Medicine;Henan Key Laboratory of Signal Transduction,Zhengzhou 450003,China)

机构地区：[1]河南中医药大学 [2]河南省方证信号传导重点实验室,郑州450003

出　　处：《天然产物研究与开发》2025年第4期602-611,共10页Natural Product Research and Development

基　　金：国家自然科学基金(82074313);2022年河南省科技研发联合基金(22301420084)。

摘　　要：探究小檗碱抑制胃癌细胞增殖、迁移及侵袭的作用机制。以AGS和HGC27胃癌细胞为研究对象,用不同浓度(10、20、30、40、50μg/mL)的小檗碱进行干预,MTT比色法检测胃癌细胞抑制率;克隆形成实验检测胃癌细胞增殖能力;流式细胞术检测胃癌细胞的周期及凋亡;划痕形成实验检测胃癌细胞迁移能力;Transwell法检测胃癌细胞的侵袭能力;转录组测序检测药物可能的作用机制;实时荧光定量聚合酶链式反应检测胃癌细胞E钙粘蛋白(E-cadherin,E-CA)、N钙粘蛋白(N-cadherin,N-CA)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、波形蛋白(vimentin,VIM)的mRNA表达;Western blot检测胃癌细胞E-CA、N-CA、α-SMA、VIM、磷酸化波形蛋白(P-vimentin,P-VIM)的蛋白表达。结果表明,与空白对照组比较,小檗碱可显著抑制胃癌细胞的细胞活性,抑制胃癌细胞的克隆形成能力(P<0.05),增加阻滞在G0/G1期的细胞数量(P<0.05),促进胃癌细胞凋亡(P<0.05),抑制胃癌细胞的迁移及侵袭能力(P<0.05)。小檗碱可降低胃癌细胞N-CA、VIM及α-SMA的mRNA和蛋白表达(P<0.05),增加E-CA的mRNA和蛋白表达(P<0.01),增加P-VIM(P<0.01)的蛋白表达。由此可知,小檗碱诱导胃癌细胞凋亡,抑制胃癌细胞的体外活性、侵袭和迁移,可能与上皮间充质转化有关。This study aims to explore the mechanism by which berberine inhibits the proliferation,migration,and invasion of gastric cancer cells(AGS and HGC27).AGS and HGC27 cells were used as the research subjects and were intervened with berberine at different concentrations(10,20,30,40,and 50μg/mL).The MTT assay was employed to detect the inhibition rate of gastric cancer cells.The colony formation assay was used to assess the proliferation ability of gastric cancer cells.Flow cytometry was utilized to determine the cell cycle and apoptosis of gastric cancer cells.The scratch formation assay was conducted to measure the migration ability of gastric cancer cells.The Transwell assay was performed to examine the invasion ability of gastric cancer cells.Transcriptome sequencing was implemented to detect the possible mechanism of action of the drug.RT-qPCR was carried out to detect the mRNA expression of E-cadherin(E-CA),N-cadherin(N-CA),α-smooth muscle actin(α-SMA),and vimentin(VIM)in gastric cancer cells.Western blot was used to detect the protein expression of E-CA,N-CA,α-SMA,VIM,and phosphorylated vimentin(P-VIM)in gastric cancer cells.Results demonstrated that compared with the blank control group,berberine significantly inhibited the cell viability of gastric cancer cells,suppressed the colony formation ability of gastric cancer cells(P<0.05),increased the number of cells arrested in the G0/G1 phase(P<0.05),promoted apoptosis of gastric cancer cells(P<0.05),and inhibited the migration and invasion abilities of gastric cancer cells(P<0.05).Berberine could reduce the mRNA and protein expression of N-CA,VIM,andα-SMA in gastric cancer cells(P<0.05),increase the mRNA and protein expression of E-CA(P<0.01),and increase the protein expression of P-VIM(P<0.01).Consequently,berberine induces apoptosis of gastric cancer cells and inhibits the in vitro activity,invasion,and migration of gastric cancer cells,which may be related to epithelial-mesenchymal transition.

关 键 词：胃癌细胞 小檗碱 迁移 侵袭 上皮间充质转化 

分 类 号：R73-34[医药卫生—肿瘤]