淫羊藿次苷Ⅱ对人胃癌细胞MKN-45和MKN-74增殖、侵袭、迁移的影响及机制  

Effect and mechanism of IcarisideⅡon proliferation,invasion,and migration of human gastric cancer cells MKN-45 and MKN-74

作　　者：孙铭 兰冰雪 谭琪琪 蒲廷伟 程树强 韦四喜 黄海 SUN Ming;LAN Bingxue;TAN Qiqi;PU Tingwei;CHENG Shuqiang;WEI Sixi;HUANG Hai(Clinical Laboratory Center,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;School of Clinical Laboratory Sciences,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学附属医院临床检验中心,贵州贵阳550004 [2]贵州医科大学医学检验学院,贵州贵阳550004

出　　处：《贵州医科大学学报》2024年第12期1729-1738,共10页Journal of Guizhou Medical University

基　　金：国家自然科学基金(82060442);贵州医科大学附属医院博士科研启动基金项目(gyfybsky-2021-39);贵州医科大学附属医院国家自然科学基金培育计划项目(gyfynsfc-2021-11);贵州医科大学国家自然科学基金培育项目(21NSFCP05)。

摘　　要：目的探讨淫羊藿次苷Ⅱ(IcarisideⅡ)对人胃癌细胞MKN-45和MKN-74增殖、侵袭、迁移的影响及机制。方法取对数生长期人胃癌细胞MKN-45和MKN-74各分为对照组(等体积二甲基亚砜)及15、30μmol/L IcarisideⅡ组,采用细胞增殖毒性实验(CCK-8)检测各组2种细胞的细胞存活率,采用Transwell实验检测各组2种细胞的侵袭和迁移能力,采用流式细胞术检测各组2种细胞的凋亡和细胞周期情况,采用集落形成实验检测各组2种细胞的克隆数量,采用Western blot检测各组2种细胞中上皮间质转化(EMT)相关蛋白[上皮钙黏蛋白(E-Cadherin)、神经钙黏蛋白(N-Cadherin)及波形蛋白(Vimentin)]和凋亡相关蛋白[B淋巴细胞瘤蛋白-2(Bcl-2)及Bcl2-相关X蛋白(Bax)]的表达。结果与对照组相比,15、30μmol/L IcarisideⅡ组MKN-45和MKN-74细胞的增殖和克隆形成能力降低,且呈时间、浓度依赖性(P<0.05);与对照组比较,15、30μmol/L IcarisideⅡ组MKN-45和MKN-74细胞的侵袭和迁移能力降低(P<0.05);与对照组比较,15、30μmol/L IcarisideⅡ组MKN-45和MKN-74细胞的总凋亡比例增加、Bcl-2表达下调及Bax蛋白表达上调(P<0.05),15、30μmol/L IcarisideⅡ组MKN-45和MKN-74细胞的G1期细胞增加(P<0.05);与对照组相比,15、30μmol/L IcarisideⅡ组MKN-45和MKN-74细胞中E-Cadherin表达上调,N-Cadherin和Vimentin表达下调(P<0.05)。结论IcarisideⅡ可影响MKN-45和MKN-74细胞的增殖、侵袭及迁移,其机制可能与其诱导细胞凋亡、阻滞细胞周期及抑制EMT有关。Objective To explore the effect and mechanism of IcarisideⅡon proliferation,invasion,and migration of human gastric cancer cells MKN-45 and MKN-74.Methods Logarithmically growing MKN-45 and MKN-74 cells were divided into control groups(with equal volumes of pure dimethyl sulfoxide)and 15,30μmol/L IcarisideⅡgroups.Cell viability was using the cell counting kit-8(CCK-8)assay.Transwell assays were performed to evaluate cell invasion and migration abilities.Flow cytometry was used to analyze the cell cycle and apoptosis in the treated cells.Colony formation assay was used to assess cell colony number.Western blot was employed to detect the expression of apoptosis-related proteins[B-cell lymphoma 2(Bcl-2)and Bcl2-associated X protein(Bax)]and EMT-related proteins[epithelial cadherin(E-Cadherin),neural cadherin(N-Cadherin),and vimentin].Results Compared with the control group,the proliferation and colony formation abilities of cells in the IcarisideⅡtreatment group were reduced in a time-and concentration-dependent manner(P<0.05).IcarisideⅡinhibited the invasion and migration abilities of gastric carcinoma cells compared with the control group(P<0.05).In the IcarisideⅡtreatment group,the total apoptosis ratio of MKN-45 and MKN-74 cells increased,Bcl-2 expression decreased,and Bax protein expression increased compared with the control group(P<0.05).IcarisideⅡtreatment increased the proportion of MKN-45 and MKN-74 cells in the G1 phase(P<0.05).Additionally,E-Cadherin expression was upregulated,while N-Cadherin and vimentin expressions were downregulated in the IcarisideⅡtreatment group compared with the control group.Conclusion IcarisideⅡaffects the proliferation,invasion,and migration of MKN-45 and MKN-74 cells,and its underlying mechanism may be related to IcarisideⅡ-induced apoptosis,cell cycle arrest,and inhibition of EMT.

关 键 词：胃肿瘤 上皮-间质转化 细胞凋亡 淫羊藿次苷Ⅱ 侵袭 迁移 

分 类 号：R735.2[医药卫生—肿瘤]