苦豆碱抑制胃癌HGC-27细胞和AGS细胞增殖的作用机制  

Mechanism of aloperine inhibiting the proliferation of HGC-27 and AGS gastric cancer cells

作　　者：锁丽娜 马海秀 易毅 苏玉东 翟少倩 赵婧 曹成珠[1] 苏占海 SUO Lina;MA Haixiu;YI Yi;SU Yudong;ZHAI Shaoqian;ZHAO Jing;CAO Chengzhu;SU Zhanhai(Medical College of Qinghai University,Xining 810001,China;Research Center for High Altitude Medicine,Qinghai University,Xining 810001,China)

机构地区：[1]青海大学医学部,西宁810001 [2]青海大学高原医学研究中心,西宁810001

出　　处：《中国高原医学与生物学杂志》2024年第4期235-245,共11页Journal of Chinese High Altitude Medicine & Biology

基　　金：国家自然科学基金项目(32360242);青海省自然科学基金项目(NO.2023-ZJ-932M)。

摘　　要：目的探讨苦豆碱(Aloperine,ALO)抑制胃癌HGC-27细胞、AGS细胞增殖的作用机制。方法用不同浓度ALO处理胃癌HGC-27细胞、AGS细胞。应用细胞毒性实验、细胞增殖实验和平板克隆实验评估增殖活性,应用划痕实验评估迁移与修复能力,应用侵袭实验检测迁移和侵袭能力,使用流式细胞术分析细胞周期变化情况,应用实时荧光定量聚合酶链反应实验测定GSPT1、CDK4、Cyclin D1和Caspase-3、Caspase-9、Bax、Bcl-2的mRNA表达水平,应用蛋白印迹实验评估GSPT1和Caspase-3、Caspase-9、Bax及Bcl-2的蛋白表达水平。结果细胞毒性、细胞增殖以及平板克隆实验结果显示,ALO抑制胃癌HGC-27细胞、AGS细胞增殖并呈现浓度依赖性:与对照组相比,用0.1 mM和0.25 mM ALO处理的HGC-27细胞、AGS细胞的增殖能力下降。划痕实验结果显示,ALO抑制了HGC-27细胞、AGS细胞的迁移修复能力。细胞侵袭检测结果显示,经ALO处理后的HGC-27细胞、AGS细胞的迁移、侵袭能力受到抑制。流式细胞术结果显示,经ALO作用后的HGC-27细胞、AGS细胞的周期发生变化。实时荧光定量聚合酶链反应实验和蛋白印迹实验结果显示,与对照组相比,经药物处理后的HGC-27细胞、AGS细胞中的GSPT1、Bcl-2、CDK4、Cyclin D1表达水平降低,Caspase-3、Caspase-9、Bax表达水平升高。结论ALO可以通过调节GSPT1和Caspase-3、Caspase-9、Bax、Bcl-2的表达水平来抑制胃癌HGC-27细胞、AGS细胞的增殖。Objective To explore the mechanism of aloperine(ALO)inhibiting the proliferation of HCC-27 and AGS gastric cancer cells.Methods HGC-27 cells and ACS cells were exposed to varying concentrations of ALO.Proliferative activities were assessed by means of CCK-8 experiments,cell proliferation experiments and plate cloning experiments.Migration and repair ability were evaluated through scratch experiments.Migration and invasion abilities were measured through transwell experiments.Flow cytometry was adopted to analyze the variations of cell cycle.The expression of mRNA levels of GSPT1,CDK4,Cyclin D1 and Caspase-3,Caspase-9,Bax,Bcl-2 were determined by using qTR-PCR.Western Blot was performed to assess the protein expression levels of CSPT1,Caspase-3,Caspase-9,Bax and Bcl-2.Results The results of CCK-8 experiments,cell proliferation experiments and plate cloning experiments showed that the proliferation of HCC-27 and AGS gastric cancer cells were inhibited by ALO in a concentration dependent manner.That is to say,compared with the control group,the proliferation ability of HGC-27 cells and AGS cells treated with 0.1 mM and 0.25 mM ALO decreased.The results of scratch experiments showed that the migration and repair ability of HGC-27 cells and AGS cells were inhibited by ALO.The results of transwell experiments showed that the migration and invasion ability of HGC-27 cells and AGS cells treated with ALO were inhibited.The results of flow cytometry showed that there were variations in the cell cycle of HGC-27 and AGS after ALO treatment.The results of qRT-PCR and Western Blot showed that compared with the control group,the expression levels of CSPT1、Bcl-2,CDK4 and Cyclin D1 in HGC-27 cells and ACS cells treated with drugs decreased,while that of Caspase-3,Caspase-9,Bax increased.Conclusion ALO may inhibit the proliferation of HCC-27 and ACS gastric cancer cells by regulating the expression levels of GSPT1,Caspase-3,Caspase-9,Bax and Bcl-2.

关 键 词：苦豆碱 抑制 HGC-27 AGS 增殖 机制 

分 类 号：R394.2[医药卫生—医学遗传学]