艾叶乙酸乙酯提取物通过LINC01606/微小RNA-26b对结直肠癌细胞增殖、凋亡的影响  

Effects of ethyl acetate extract of Folium Artemisiae Argyi on the proliferation and apoptosis of colorectal cancer cells through LINC01606/miR-26b

作　　者：蒋学军[1] 杨勇[1] 庹磊 吉祖进 雷新益 JIANG Xuejun;YANG Yong;TUO Lei;JI Zujin;LEI Xinyi(Department of Colorectal and Anal Surgery,Dongfeng General Hospital of Sinopharm,Hubei Medical College,Shiyan,Hubei 442000,China)

机构地区：[1]湖北医药学院附属国药东风总医院结直肠肛门外科,湖北十堰442000

出　　处：《安徽医药》2022年第9期1719-1723,共5页Anhui Medical and Pharmaceutical Journal

摘　　要：目的研究艾叶乙酸乙酯提取物对结直肠癌SW1116细胞增殖和凋亡的影响及其潜在的分子机制。方法2018年6月至2019年11月,用12.5 mg/L、25 mg/L、50 mg/L的艾叶乙酸乙酯提取物处理结直肠癌细胞SW1116,分别称为实验1、2、3组,人胆囊收缩素/缩胆囊素八肽(CCK-8)法、流式细胞术和克隆形成实验检测SW1116细胞存活率、凋亡率、细胞周期和克隆形成能力,蛋白质印迹法检测周期素依赖激酶抑制剂p21(P21)和胱天蛋白酶-3(caspase-3)蛋白表达水平,实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测细胞中LINC01606和微小RNA(miR)-26b的转录水平,双荧光素酶报告系统验证LINC01606和miR-26b的关系。结果实验1、2、3组的SW1116细胞克隆形成数(101±9、79±8、56±6)、存活率[(81.57±8.02)%、(68.17±6.69)%、(48.26±4.91)%]、S期细胞百分比[(31.98±2.39)%、(26.10±2.21)%、(23.21±2.04)%]及细胞中LINC01606含量(0.76±0.05、0.51±0.05、0.34±0.04)逐渐降低(P<0.05),P21(0.34±0.03、0.47±0.04、0.67±0.05)、caspase-3(0.26±0.02、0.38±0.03、0.59±0.04)蛋白表达量、miR-26b含量(1.26±0.09、1.41±0.10、1.69±0.12)、细胞凋亡率[(13.37±1.21)%、(17.94±1.47)%、(24.36±1.69)%]和G1期细胞百分比[(32.17±2.41)%、(37.04±2.75)%、(41.20±3.24)%]均逐渐升高(P<0.05),呈显著剂量依赖趋势,剂量越高效果越显著;过表达LINC01606和抑制miR-26b均可提高细胞克隆形成数、存活率和S期细胞百分比,降低P21、caspase-3蛋白表达量、细胞凋亡率及G1期细胞百分比;LINC01606靶向调控miR-26b。结论艾叶乙酸乙酯提取物通过LINC01606/miR-26b途径抑制SW1116细胞的增殖,促进细胞凋亡。艾叶乙酸乙酯提取物对结直肠癌具有潜在治疗作用。Objective To study the effect of ethyl acetate extract of Folium Artemisiae Argyi on the proliferation and apoptosis of colorectal cancer SW1116 cells and its underlying molecular mechanism.Methods From June 2018 to November 2019,SW1116 colorectal cancer cells were treated with 12.5 mg/L,25 mg/L and 50 mg/L ethyl acetate extracts from Folium Artemisiae Argyi,referred to as experiments 1,2,and 3,respectively.The human cholecystokinin/cholecystokinin octapeptide(CCK-8)method,flow cytometry and colony formation assays were used to detect the survival rate,apoptosis rate,cell cycle and colony formation ability of SW1116 cells.Western blotting was used to detect the levels of P21 and caspase-3 proteins,quantitative real-time polymerase chain reaction(qRT‒PCR)was used to detect the levels of long noncoding RNA LINC01606 and microRNA(miR)-26b,and a dual-luciferase reporter system was used to verify the relationship between LINC01606 and miR-26b.Results The number of colonies of SW1116 cells in ex-perimental groups 1,2 and 3(101±9,79±8,56±6),survival rate[(81.57±8.02)%,(68.17±6.69)%,(48.26±4.91)%],percentage of cells in S phase[(31.98±2.39)%,(26.10±2.21)%,(23.21±2.04)%],and LINC01606 content in cells(0.76±0.05,0.51±0.05,0.34±0.04)gradu-ally decreased(P<0.05).The P21(0.34±0.03,0.47±0.04,0.67±0.05),caspase 3(0.26±0.02,0.38±0.03,0.59±0.04)proteins,miR-26b content(1.26±0.09,1.41±0.10,1.69±0.12),apoptosis rate[(13.37±1.21)%,(17.94±1.47)%,(24.36±1.69)%]and percentage of cells in G1-phase[(32.17±2.41)%,(37.04±2.75)%,(41.20±3.24)%]gradually increased(P<0.05),showing a significant dose-dependent trend,and the higher the dose,the more significant the effect.Overexpression of LINC01606 and inhibition of miR-26b can both increase the number of cell clones and survival.The percentage of cells in S phase and the P21 and caspase-3 protein expression,apoptosis rate and percentage of cells in G1 phase were reduced.LINC01606 targets and regulates miR-26b.Conclusion The ethyl acetate extract of Fo⁃lium Artemi

关 键 词：艾叶乙酸乙酯提取物 结直肠肿瘤 长链非编码RNA LINC01606 增殖 凋亡 

分 类 号：R285[医药卫生—中药学]