摘要
目的探讨信号转导和转录激活因子3(STAT3)抑制剂盐酸萘替芬(stattic)对小鼠结肠癌CT26细胞增殖和凋亡的影响和作用机制。方法采用0μmol/L、1μmol/L、5μmol/L、10μmol/L stattic溶液处理小鼠结肠癌CT26细胞,通过CCK-8实验、细胞克隆形成实验、细胞划痕实验、Transwell侵袭实验以及流式细胞术检测细胞活力、增殖、迁移、侵袭、周期和凋亡情况;利用Western blot法检测stattic对小鼠结肠癌细胞磷酸化STAT3(p-STAT3)表达的影响;通过实时荧光定量多聚核苷酶链式反应(RT-qPCR)检测stattic作用后CT26细胞B淋巴细胞瘤-2(Bcl-2)和人跨膜受体蛋白Notch-1(Notch-1)的表达。结果与0μmol/L组相比,stattic溶液组CT26细胞的活力及增殖能力降低(P<0.001)、迁移率和侵袭率降低(P<0.001),细胞凋亡率随浓度增加而增加(P<0.0001);stattic能将CT26细胞周期阻断于G1期,进而阻止CT26细胞的增殖;Western blot结果显示stattic抑制CT26细胞p-STAT3的表达(P<0.05);RT-qPCR检测结果表明stattic下调CT26细胞Bcl-2和Notch1的表达(P<0.05)。结论stattic通过阻断STAT3信号,抑制p-STAT3蛋白的表达,下调下游抗凋亡分子Bcl-2和Notch1信号分子的表达从而抑制CT26细胞增殖促进细胞凋亡。
Objective To investigate the mechanism and effect of signal transduction and signal transducer and activator of transcription 3(STAT3)inhibitor naphthoprofen hydrochloride(stattic)on the proliferation and apoptosis of mouse colon cancer CT26 cells.Methods Mouse colon cancer CT26 cells were treated with control group stattic solution of 0,1,5,and 10μmol/L.Cell viability,proliferation,migration,invasion,cycle and apoptosis were investigated by CCK-8 assay,cell clonogenesis assay,cell scratch assay,Transwell assay and flow cytometry.The effect of stattic on phosphorylated STAT3(p-STAT3)expression in mouse colon cancer cells was detected by Western blot.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression of B lymphoblastoma-2(Bcl-2)and human transmembrane receptor Notch-1(Notch-1)in CT26 cells after stattic treatment.Results Compared with the 0μmol/L control group,the viability and proliferation of CT26 cells of static solution group decreased(P<0.001),the migration and invasion rate decreased(P<0.001),and the apoptosis rate increased with the increase of CT26 concentration(P<0.0001).stattic could block the cell cycle of CT26 in G1 phase,thus preventing the proliferation of CT26 cells.Western blot results showed that stattic inhibited the expression of p-STAT3 in CT26 cells(P<0.05).The results of RT-qPCR showed that stattic down-regulated the expression of Bcl-2 and Notch1 in CT26 cells(P<0.05).Conclusion stattic inhibited the expression of p-STAT3 protein and down-regulated expression of down-stream anti-apoptotic Bcl-2 and Notch1 signaling molecules by blocking STAT3 signal,thus inhibiting the proliferation of CT26 cells and promoting cell apoptosis.
作者
张瑾宬
缪心怡
操蓉
黎敏
张儒雅
刘丽娜
ZHANG Jincheng;MIAO Xinyi;CAO Rong;LI Min;ZHANG Ruya;LIU Li'na(School of Biology and Engineering&School of Modern Medicine and Health Care Industry,Guizhou Medical University,Guiyang 550025,Guizhou,China;School of Basic Medical Science,Guizhou Medical University,Guiyang 550025,Guizhou,China)
出处
《贵州医科大学学报》
CAS
2024年第4期522-528,560,共8页
Journal of Guizhou Medical University
基金
贵州省科技计划项目(黔科合支撑〔2020〕4Y233)
国家级大学生创新创业训练计划项目(202110660008)。