环巴胺对子宫内膜癌HEC-1A细胞迁移、周期及自噬的影响  

Effect of cyclopamine on migration,cell cycle and autophagy of endometrial carcinoma HEC-1A cells

作　　者：张萍萍[1] 李雪[2] 刘元林[2] 张思 刘朝晖 童英[1] 张毅[2] 

机构地区：[1]中国人民解放军空军总医院妇产科,北京100142 [2]军事医学科学院基础医学研究所细胞生物学研究室,北京100850 [3]通州新华医院妇产科,北京101100 [4]大兴妇幼保健院,北京102600

出　　处：《中国生物制品学杂志》2017年第5期494-499,共6页Chinese Journal of Biologicals

基　　金：国家自然科学基金(31070996)

摘　　要：目的探讨环巴胺(cyclopamine,CYP)对子宫内膜癌HEC-1A细胞迁移、周期及自噬的影响。方法分别用0、5、10、20、40μmol/L CYP处理HEC-1A细胞24和/或48 h,瑞氏吉姆萨染色后观察细胞形态改变,划痕试验检测细胞迁移能力,流式细胞术检测细胞周期及自噬微管相关蛋白轻链3B(light chain 3B,LC3B)的表达,Western blot法检测LC3-B蛋白表达水平。结果随着CYP浓度的增加,HEC-1A细胞数目减少,细胞密度逐渐降低,出现空泡化、核碎裂等现象逐渐加重;HEC-1A细胞的迁移率显著下降(P<0.05);HEC-1A细胞处于G0/G1期的比例显著增加(P<0.05);表达LC3B蛋白的HEC-1A细胞百分比显著上升(P<0.05),且HEC-1A细胞中的LC3B蛋白表达水明显增高(P<0.05)。结论 CYP可阻滞子宫内膜癌HEC-1A细胞周期运转,引发细胞自噬,从而抑制细胞存活及迁移。Objective To investigate the effect of cyclopamine（CYP） on the migration, cell cycle and autophagy of endometrial carcinoma HEC-1A cells. Methods HEC-1A cells were treated with CYP at concentrations of 0, 5, 10, 20 and 40 μmol/L for 24 and/or 48 h, and observed for morphology by Giemsa staining, tested for migration ability by wound healing test, for cell cycle and expression level of autophagy microtubule associated protein light chain 3B（LC3B）by flow cytometry, and for expression level of LC3-B protein by Western blot. Results Along with the increasing CYP concentration, HEC-1A cells decreased in quantity and density, while vacuolization and karyorrhexis were aggravated.The migration rate of HEC-1A cells decreased significantly（P〈0. 05）, while the percentage of cells at G0/G1 phases, the percentage of HEC-1A cells positive for LC3 B protein and the expression level of LC3 B protein increased significantly（each P〈0. 05）. Conclusion CYP arrested the cell cycle of HEC-1A cells and induced the cell autophagy, thus inhibited the survival and migration of the cells.

关 键 词：环巴胺 子宫内膜癌HEC-1A细胞 细胞周期 细胞迁移 自噬 

分 类 号：R737.33[医药卫生—肿瘤] R979.12[医药卫生—临床医学]