氧化苦参碱诱导胃癌BGC-823细胞发生caspase-12依赖性内质网应激凋亡的机制研究  

Mechanism of caspase-12-dependent endoplasmic reticulum stress apoptosis induced by oxymatrine in gastric cancer BGC-823 cells

作　　者：房伟[1] 马世勋[1] 马云涛[1] 詹渭鹏[1] 李渊[1] 李一平[1] 狐鸣[1] FANG Wei;MA Shixun;MA Yuntao;ZHAN Weipeng;LI Yuan;LI Yiping;HU Ming(Department of General Surgery 1,Gansu Provincial Hospital,Lanzhou 730000,P.R.China)

机构地区：[1]甘肃省人民医院普外一科,兰州730000

出　　处：《中国普外基础与临床杂志》2019年第4期399-404,共6页Chinese Journal of Bases and Clinics In General Surgery

基　　金：甘肃省自然科学基金项目(项目编号:1508RJZA043)

摘　　要：目的研究氧化苦参碱对胃癌BGC-823细胞增殖和凋亡的影响,并探讨内质网应激参与氧化苦参碱诱导胃癌细胞的凋亡情况并阐明其机制。方法将对数生长期的胃癌BGC-823细胞分为对照组、单药组(用10、30、60和90μmol/L浓度的氧化苦参碱处理)和联合组(各浓度的氧化苦参碱联合2μmol/L内质网应激抑制剂salubrinal处理)。用MTT法观察氧化苦参碱对胃癌BGC-823细胞生长的抑制作用,用流式细胞仪分析氧化苦参碱诱导胃癌BGC-823细胞凋亡及其对细胞周期的影响,用Western blot和RT-PCR法分别检测内质网应激标志分子GRP78/Bip和内质网应激介导凋亡分子caspase-12蛋白和基因的表达情况。结果 (1)与对照组比较,氧化苦参碱能明显抑制胃癌BGC-823细胞生长且呈浓度-时间依赖性(P<0.05),其IC_(50)(48 h)值为(59.5±0.5)μmol/L;与相应浓度单药组比较,30、60和90μmol/L氧化苦参碱联合作用对细胞的抑制作用明显减弱(P<0.05)。(2)与对照组比较,随着氧化苦参碱浓度的增加,G2/M期细胞数和细胞凋亡率呈增加(或增高)趋势(P<0.05);与60μmol/L氧化苦参碱单药组比较,60μmol/L氧化苦参碱联合组处理胃癌BGC-823细胞48 h后明显降低细胞凋亡率和减少G_2/M期细胞数(P<0.05)。(3)与对照组比较,随着氧化苦参碱浓度的增加(除外10μmol/L氧化苦参碱组caspase-12蛋白及基因),单药组GRP78/Bip和caspase-12蛋白及基因表达水平呈明显增高趋势(P<0.05);与相应浓度单药组比较,60及90μmol/L氧化苦参碱联合作用时胃癌BGC-823细胞中GRP78/Bip和caspase-12蛋白及基因表达水平均明显降低(P<0.05)。结论氧化苦参碱对人胃癌BGC-823细胞有明显的生长抑制作用,该作用可能与caspase-12依赖性诱导凋亡及上调GRP78/Bip表达水平有关,该结论需要进一步实验验证。Objective To study effects of oxymatrine on proliferation and apoptosis of gastric cancer cell line BGC-823 and explore role of endoplasmic reticulum stress in apoptosis of gastric cancer cells induced by oxymatrine and elucidate its mechanism.Methods The gastric cancer BGC-823 cells at the logarithmic phase were divided into a control group,oxymatrine alone group(oxymatrine at 10,30,60 and 90|imol/L concentrations),and combination group(oxymatrine at various concentrations combined with 2μmol/L endoplasmic reticulum stress inhib让or salubrinal).The MTT assay was used to observe the inhibitory effect of the oxymatrine on the growth of BGC-823 cells.The flow cytometry was used to analyze the apoptosis and cell cycle.The Western blot and RT-PCR methods were used to detect the expressions of GRP78/Bip and the caspase-12 protein and gene,respectively.Results①Compared with the control group,the oxymatrine could significantly inhib让the growth of gastric cancer BGC-823 cells in a concentration-time dependent manner(P<0.05)and its IC50(48 h)value was(59.5±0.5)pmol/L.The inhib让ory effect of the combination group of 30,60,and 90 pmol/L oxymatrine was significantly weakened as compared with the the corresponding oxymatrine alone group(P<0.05).②The oxymatrine could significantly induce the apoptosis and arrest the G2/M phase in the gastric cancer BGC-823 cells in a concentration-dependent manner(P<0.05).The combination group of 60μmol/L oxymatrine could significantly decreased the apoptosis rate and the number of cells in the G2/M phase in the gastric cancer BGC-823 cells after treating 48 h(P<0.05).③The protein and gene levels of GRP78/Bip and caspase-12 showed significant increases with the increase of oxymatrine concentrations in the oxymatrine alone group(except the protein and gene levels of caspase-12 at 10μmol/L oxymatrine)as compared with the control group(P<0.05),which in the combination group of 60 pmol/L and 90 pmol/L oxymatrines were significantly decreased as compared with the corresponding oxymat

关 键 词：氧化苦参碱 胃肿瘤 凋亡 内质网应激 

分 类 号：R735.2[医药卫生—肿瘤]