鬼臼苦素对人结直肠癌HCT-15细胞增殖和周期的影响及其作用机制  

Effects of picropodophyllin on proliferation and cell cycle of human colorectal cancer HCT-15 cells and its mechanism

作　　者：周丽丽 王瑞婷[2] 赵杨[1] 薛晶[3] 刘赛璇 申兴斌[1] ZHOU Li-li;WANG Rui-ting;ZHAO Yang;XUE Jing;LIU Sai-xuan;SHEN Xing-bin(Department of Pathology,the Affiliated Hospital of Chengde Medical University,Hebei Chengde 067000,China;Department of Pharmacology,Chengde Medical University,Hebei Chengde 067000,China;Experiment Center for Morphology,Chengde Medical University,Hebei Chengde 067000,China)

机构地区：[1]承德医学院附属医院病理科,河北承德067000 [2]承德医学院药理学教研室,河北承德067000 [3]承德医学院形态学实验中心,河北承德067000

出　　处：《解剖学报》2020年第2期233-238,共6页Acta Anatomica Sinica

摘　　要：目的探讨鬼臼苦素(PPP)对人结直肠癌细胞系HCT-15细胞增殖和周期的影响及其作用机制。方法不同浓度PPP处理HCT-15细胞,采用细胞计数试剂盒8(CCK-8)法检测HCT-15细胞的增殖活性;倒置显微镜下观察HCT-15细胞的形态变化;流式细胞术检测HCT-15细胞周期的改变;Western blotting检测增殖细胞核抗原(PCNA)及细胞周期蛋白D1(cyclin D1)的蛋白表达水平。结果CCK-8法结果显示,PPP能显著抑制HCT-15细胞的增殖活性,并呈剂量-时间依懒性;倒置显微镜下观察发现,细胞失去原有形态回缩为圆形,折光性降低,活细胞数量明显减少;流式细胞术结果显示,G2/M期细胞比例显著升高,G0/G1期和S期细胞比例明显降低;Western blotting结果显示,PCNA、细胞周期蛋白D1的表达水平明显降低。结论PPP可显著抑制人结直肠癌HCT-15细胞的增殖,其作用机制可能是通过下调PCNA、cyclin D1的表达水平,进而将细胞阻滞于G2/M期发挥抑制作用。Objective To investigate the effect of picropodophyllin(PPP)on proliferation and cell cycle of human colorectal cancer cell line HCT-15 and to clarify the related mechanism.Methods Different concentrations of PPP were used to treat HCT-15 cells,the proliferative activity of HCT-15 cells was detected by cell courting kit-8(CCK-8).Morphological changes of HCT-15 cells were observed under inverted microscope.Flow cytometry was used to detect the changes of HCT-15 cell cycle and Western blotting was used to detect the expression of proliferating cell nuclear antigen(PCNA)and cell cyclin D1.Results The result of CCK-8 assay showed that PPP inhibited the proliferation of HCT-15 cells in a dose-time-dependent manner.Under the inverted microscope,it was found that the HCT-15 cells lost their original shape and became round,the refractive index decreased,and the number of living cells decreased significantly.Flow cytometry showed that the proportion of G2/M cells increased significantly,and the proportion of G0/G1 phase and S phase cells decreased significantly.Western Blot result showed that the expression of PCNA,cyclin D1 protein was significantly decreased.Conclusion PPP significantly inhibited the proliferation of human colorectal cancer HCT-15 cells,suggesting that the mechanism might be to block the cells in G2/M phase by down-regulating the expression of PCNA,cyclin D1 protein.

关 键 词：鬼臼苦素 结直肠癌HCT-15细胞 流式细胞术 免疫印迹法 人 

分 类 号：R735.3[医药卫生—肿瘤]