利拉鲁肽对结肠癌细胞增殖、凋亡及转录活化因子6通路的影响  

The influences of liraglutide on proliferation,apoptosis,and ATF6 pathway of colon cancer cells

作　　者：智会[1] 张志永[2] ZHI Hui;ZHANG Zhiyong(Department of Anorectal Surgery,Zhoukou Central Hospital,Zhoukou,Henan 466000,China;Department of Anorectal Surgery,The First Affiliated Hospital of Zhengzhou University,Zhengzhou,Henan 450000,China)

机构地区：[1]周口市中心医院肛肠外科,河南周口466000 [2]郑州大学第一附属医院肛肠外科,河南郑州450000

出　　处：《安徽医药》2020年第3期438-442,I0003,共6页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探究利拉鲁肽对结肠癌细胞增殖、凋亡的影响及对转录活化因子6(ATF6)通路的影响。方法按照利拉鲁肽浓度0、10、100、1000 nM培养人结肠癌HCT116细胞24、48、72 h,人胆囊收缩素/缩胆囊素八肽(CCK⁃8)法检测细胞增殖情况,流式细胞仪检测细胞周期、凋亡情况,蛋白质印迹法(Western Blot)检测作用后细胞中21KD蛋白(Bax)、半胱胺酸蛋白酶⁃3(Caspase⁃3)及ATF6通路蛋白ATF6 p50、CCAAT⁃增强子结合蛋白(C/EBP)同源蛋白(CHOP)的表达水平。结果与对照组相比,利拉鲁肽各浓度间增殖率均有显著变化(P<0.05),同一时间点随着利拉鲁肽浓度的增加,0、10、100、1000 nM各组细胞的增殖率分别为(97.16±35.67)%、(81.69±33.37)%、(76.12±30.23)%、(70.65±28.89)%;(86.17±33.98)%、(58.68±26.77)%、(39.74±10.67)%、(30.17±9.24)%;(83.82±31.19)%、(38.77±10.19)%、(22.98±6.78)%、(16.63±6.12)%,均呈下降趋势。同一浓度随着时间推移,24、48、72 h各时间点细胞增殖率分别为(97.16±35.67)%、(86.17±33.98)%、(83.82±31.19)%;(81.69±33.37)%、(58.68±26.77)%、(38.77±10.19)%;(76.12±30.23)%、(39.74±10.67)%、(22.98±6.78)%;(70.65±28.89)%、(30.17±9.24)%、(16.63±6.12)%。不同浓度利拉鲁肽作用于HCT116细胞72 h后,10、100、1000 nM浓度的利拉鲁肽组的G2/M期细胞比例较0 nM组显著减少(P<0.05),且随着浓度的增加G2/M期细胞呈现下降趋势。0、10、100、1000 nM利拉鲁肽作用下细胞凋亡率分别为(9.06±0.98)%、(10.45±1.12)%、(13.89±1.54)%、(56.08±14.33)%,依次升高(P<0.05)。与0 nM组比较,10、100、1000 nM浓度的利拉鲁肽组细胞的凋亡率依次显著升高(P<0.05);与10 nM组比较,100、1000 nM组凋亡率依次显著升高(P<0.05);与100 nM组比较,1000 nM组凋亡率显著升高(P<0.05)。蛋白质印迹法结果显示,10、100、1000 nM利拉鲁肽作用后细胞中Bax蛋白表达量分别为(0.69±0.13)、(0.93±0.24)、(1.19±0.25),显著升高(P<0.05),1000 nM组Bax蛋白表达�Objective To investigate the influences of liraglutide on proliferation,apoptosis and ATF6 pathway of colon cancer cells.Methods Human colon cancer HCT116 cells were cultured at liraglutide concentrations of 0,10,100,and 1000 nM for 24,48,and 72 h.Cell proliferation was measured by the human cholecystokinin/cholecystokinin octapeptide(CCK⁃8)method.Cell cycle and apoptosis were detected by cell cytometry,21KD protein(Bax),caspase⁃3 and activated transcription factor 6(ATF6)pathway were detected by Western Blot Protein ATF6 p50,CCAAT⁃enhancer binding protein(C/EBP)homologous protein(CHOP)expres⁃sion levels.Results Compared with the control group,the proliferation rate of all concentrations of liraglutide significantly changed(P<0.05).At the same time,with the increase of liraglutide concentration,the cell proliferation rate of 0,10,100,and 1000 nM group was(97.16±35.67)%,(81.69±33.37)%,(76.12±30.23)%,(70.65±28.89)%;(86.17±33.98)%,(58.68±26.77)%,(39.74±10.67)%,(30.17±9.24)%;(83.82±31.19)%,(38.77±10.19)%,(22.98±6.78)%,and(16.63±6.12)%,all of which exhibit a downward trend.With the same concentration over time,the cell proliferation rate at 24,48,and 72 h point was(97.16±35.67)%,(86.17±33.98)%,(83.82±31.19)%;(81.69±33.37)%,(58.68±26.77)%,(38.77±10.19)%;(76.12±30.23)%,(39.74±10.67)%,(22.98±6.78)%;(70.65±28.89)%,(30.17±9.24)%,(16.63±6.12)%.After 72 h of treatment with different concentrations of liraglutide on HCT116 cells,the proportion of G2/M phase cells in 10,100,and 1000 nM liraglutide groups was significantly lower than that in 0 nM group(P<0.05),and G2/M cells showed a decreasing trend with the increase of drug concentration.The apoptosis rate increased under the action of 0,10,100,and 1,000 nM liraglutide were(9.06±0.98)%,(10.45±1.12)%,(13.89±1.54)%,(56.08±14.33)%,respectively(P<0.05).Compared with the 0 nM group,the apoptosis rate of cells in the liraglutide group at the concentrations of 10,100,and 1000 nM significantly increased sequentially(P<0.05).Compared with the 10 nM

关 键 词：结肠肿瘤 基因 ras CCAAT增强子结合蛋白类 半胱氨酸天冬氨酸蛋白酶 原癌基因蛋白质c⁃bcl⁃2 细胞增殖 细胞凋亡 利拉鲁肽 转录活化因子6通路 

分 类 号：R96[医药卫生—药理学]