参麦注射液对胃癌细胞化疗耐药的影响及其机制  

Effect of Shenmai injection on chemotherapy resistance of gastric cancer cells and underlying mechanism

作　　者：李步卓 冯联忠[1] 何春华[1] 胡毕文[1] Li Buzhuo;Feng Lianzhong;He Chunhua;Hu Biwen(Department of Anorectal Surgery,the Second Affiliated Hospital of Jiaxing University,Jiaxing 314000,China)

机构地区：[1]嘉兴学院附属第二医院胃肠外科,嘉兴314000

出　　处：《中华实验外科杂志》2024年第1期57-61,共5页Chinese Journal of Experimental Surgery

基　　金：嘉兴市科技计划项目(2022AD30034);嘉兴市秀洲区科技计划项目(2022C001)。

摘　　要：目的探讨参麦注射液通过调控微小RNA-140-3p(miR-140-3p)对胃癌细胞多药耐药的影响与机制。方法建立并培养人胃癌耐药细胞株HGC-27/DCF(5-Fu、顺铂和多西他赛)细胞(购自美国细胞培养物收藏中心),设空白对照组(DCF+生理盐水)、阴性对照组(DCF+miR-140-3p inhibitor NC+生理盐水)、参麦组(DCF+miR-140-3p inhibitor NC+参麦注射液)、miR-140-3p抑制剂组(DCF+miR-140-3p inhibitor+生理盐水)、参麦+miR-140-3p抑制剂组(DCF+miR-140-3p inhibitor+参麦注射液)。使用荧光定量RT-qPCR和蛋白质印迹法(Western blot)实验测定miR-140-3p、程序性死亡因子-1(PD-1)、程序性死亡因子配体1(PD-L1)的表达水平;使用细胞计数试剂盒(CCK-8)测定HGC-27/DCF细胞的增殖水平;经细胞经膜联蛋白-V(Annexin V)与碘化丙锭(PI)染色测定细胞凋亡率;细胞侵袭实验(Transwell)实验测定细胞侵袭能力。使用单因素方差分析进行组间比较。结果GES-1、HGC-27、HGC-27/DCF细胞的miR-140-3p表达水平分别为3.02±0.67、1.04±0.17及0.26±0.08(F=33.800,P<0.05)、PD-1表达水平分别为0.21±0.04、0.30±0.05、0.59±0.08(F=33.800,P<0.05),PD-L1表达水平分别为0.09±0.02、0.16±0.03、0.39±0.05(F=58.355,P<0.05);事后两两比较结果显示,miR-140-3p在HGC-27/DCF细胞中的表达水平高于GES-1和DCF细胞,PD-1和PD-L1在HGC-27/DCF细胞中的表达水平低于GES-1和HGC-27细胞,组间差异有统计学意义(P<0.05)。空白对照组、阴性对照组、miR-140-3p抑制剂组、参麦组、参麦+miR-140-3p抑制剂组的细胞增殖活力分别为0.47±0.03、0.48±0.02、0.30±0.04、0.68±0.11、0.45±0.04(F=16.565,P<0.05)、细胞凋亡率分别为(8.52±0.75)、(9.02±0.66)、(21.52±4.25)、(3.88±0.55)、(15.63±3.02)%(F=25.014,P<0.05);事后两两比较结果显示,空白对照组的细胞增殖活力高于miR-140-3p抑制剂组但低于参麦组,参麦+miR-140-3p抑制剂组的细胞增殖活力高于miR-140-3p抑制剂组但低于参麦组,组间差异有统计�Objective To explore the underlying mechanism of how shenmai injection regulate the multidrug resistance of gastric cancer cells by regulating microRNA-140-3P(miR-140-3P).Methods The human gastric cancer multidrug-resistant cell line HGC-27/DCF(5-Fu,cisplatin and docetaxel)cells(purchased from the Cell Culture Collection,USA)were established and cultured.These cells were divided into following groups:control group(DCF+saline),negative control group(DCF+miR-140-3p inhibitor NC+saline),shenmai injection group(DCF+miR-140-3p inhibitor NC+shenmai injection injection),miR-140-3p inhibitor group(DCF+miR-140-3p inhibitor+saline),and shenmai injection+miR-140-3p inhibitor group(DCF+miR-140-3p inhibitor+shenmai injection injection).The levels of miR-140-3p,PD-1,PD-L1 were determined by RT-qPCR and western blot assays.The proliferation levels of HGC-27/DCF cells were determined by cell proliferation and toxicity assay kit(CCK-8).The apoptosis rate of HGC-27/DCF cells were determined by annexin V and propidium iodide(PI).The cell invasion assay(Transwell)assay was adopted to determine the cell invasion ability.One-way analysis of variance(ANOVA)was applied for comparison between groups.Results The expression levels of miR-140-3p in GES-1,HGC-27,and HGC-27/DCF cells were 3.02±0.67,1.04±0.17,and 0.26±0.08,respectively(F=33.800,P<0.05),the expression levels of PD-1 were 0.21±0.04,0.30±0.05,0.59±0.08,respectively(F=33.800,P<0.05),and expression levels of PD-L1 were 0.09±0.02,0.16±0.03,0.39±0.05(F=58.355,P<0.05),respectively;post hoc two-by-two comparisons showed that miR-140-3p expression levels were higher than those in HGC-27/DCF cells than in GES-1 and DCF cells,PD-1 and PD-L1 were expressed at lower levels in HGC-27/DCF cells than those in GES-1 and HGC-27 cells,and the differences between the groups were statistically different(P<0.05).The cell proliferation viability in the control group,negative control group,miR-140-3p inhibitor group,shenmai injection group,and shenmai injection+miR-140-3p inhibitor group were

关 键 词：参麦注射液 胃癌 多药耐药 

分 类 号：R285[医药卫生—中药学]