摘要
目的研究青蒿琥酯(ART)对人结肠癌Lovo细胞增殖的影响和可能的分子机制。方法采用噻唑蓝实验(MTT)检测ART对Lovo细胞的增殖抑制作用,用流式细胞术和电镜检测ART对细胞凋亡的影响,荧光素酶报告质粒检测ART对Wnt/β-catenin通路中TCF4/LEF转录活性的影响,Western blot检测ART对细胞内β-catenin、GSK-3β、cMyc以及凋亡相关蛋白caspase-3表达的影响。结果与对照组相比,ART能抑制Lovo细胞增殖,72 h、320μmol·L^(-1)ART的细胞抑制率高达(78.99±1.95)%(F=898.301,P=0.000)。ART能诱导细胞凋亡,24 h、160μmol·L^(-1)ART的细胞早期凋亡率达到(19.00±0.05)%,同时电镜观察到细胞凋亡的形态学表现。ART能降低TCF4/LEF报告质粒的荧光素酶活性,24 h、160μmol·L^(-1)ART可使TCF4/LEF报告质粒的荧光素酶活性降至(0.36±0.30)%(F=470.954,P<0.01);ART处理48 h后,GSK-3β和caspase-3呈浓度依赖性升高(P<0.01),而β-catenin和c-Myc蛋白水平呈药物浓度依赖性降低(P<0.01)。结论青蒿琥酯能明显抑制结肠癌Lovo细胞增殖,并促进其凋亡,可能与抑制Wnt/β-catenin信号通路活化有关。
Aim To investigate the promoting apoptosis effect of artesunate( ART) on human colon cancer Lovo cells and its mechanisms. Methods MTT assay was performed to determine the anti-proliferative effect of artesunate. Flow cytometry assay and electron micros-copy( EM) were used to evaluate the apoptotic effect of artesunate. Luciferase reporter assay was introduced to measure the activation of Wnt/β-catenin pathway. Western blot was used to detect the pathway-related protein levels of β-catenin, GSK-3β,c-Myc and apop-tosis-related protein level of casepase-3 . Results Compared with the control group, the inhibitory rate of cell proliferation at 72 h and 320 μmol·L-1 ART was (78. 99 ± 1. 95 )% ( F =898. 301, P =0. 000 ); the cell apoptotic rate at 24 h and 160 μmol · L-1 ART was(19. 00 ± 0. 05)% and morphological signs of cell apoptosis were found by EM;the transcriptional activi-ty of TCF4/LEF at 24 h and 160 μmol·L-1 ART was (0. 36 ± 0. 30)%(F =470. 954,P <0. 01); the ex-pressions of caspase-3 and GSK-3β were significantly increased, whileβ-catenin and c-Myc were significant-ly decreased when treated with different concentrations of ART for 48 h ( P <0. 01 ) . Conclusion ART may significantly inhibit proliferation and promote apoptosis of Lovo cells probably by inactivating Wnt/β-catenin pathway.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2016年第5期707-711,共5页
Chinese Pharmacological Bulletin
基金
河北省高等学校科学技术研究青年基金项目(No QN20131078)
河北省高等学校科学技术研究重点项目(No ZD20131004)
