自噬溶酶体抑制剂氯化铵促进维生素K3诱导人宫颈癌HeLa细胞凋亡的作用  

Role of autophagolysosomal inhibitor NH4Cl in apoptosis of human cervical cancer HeLa cells induced by vitamin K3

作　　者：于春艳[1] 刘希[2] 张钰[3] 苏静[3] 刘玉和[1] 

机构地区：[1]北华大学基础医学院病理学教研室,吉林吉林132013 [2]北华大学化学与生物学院,吉林吉林132013 [3]吉林大学白求恩医学院病理生理学教研室,吉林长春130021

出　　处：《中国病理生理杂志》2013年第7期1197-1201,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81202552;No.81272876);吉林省教育厅资助课题(No.吉教科合字2011第117号;No.吉教科合字2012第394号);吉林省科技厅自然科学基金资助课题(No.201215103)

摘　　要：目的:探讨自噬溶酶体抑制剂氯化铵(NH4Cl)在维生素K3(VK3)诱导的HeLa细胞凋亡过程中的作用。方法:实验分为4组:正常对照组、30μmol/L VK3作用组、10 mmol/L NH4Cl作用组和30μmol/L VK3+10mmol/L NH4Cl联合作用组。MTT法检测细胞存活率;吖啶橙(AO)染色观察溶酶体膜通透性变化,单丹磺酰尸胺(MDC)染色观察自噬泡的变化,BCECF-AM染色流式细胞术检测细胞浆pH变化,Hoechst 33342染色检测细胞核染色质凝聚状态及细胞凋亡率。结果:与对照组相比,单独应用NH4Cl对HeLa细胞无明显影响。VK3作用组He-La细胞存活率降低,AO染色荧光没有变化,细胞内出现自噬泡,细胞浆呈酸性,细胞核染色质凝聚,细胞发生凋亡。NH4Cl与VK3联合作用于HeLa细胞时,与单独应用VK3组相比,溶酶体通透性增加,细胞内自噬泡更加聚集,细胞浆酸性增强,细胞凋亡率明显增加。结论:自噬溶酶体抑制剂NH4Cl促进了VK3诱导的HeLa细胞凋亡,表明自噬可能在VK3诱导的HeLa细胞损伤过程中起到保护作用。AIM： To investigate the role of autophagolysosomal inhibitor ammonium chloride（NH4Cl） in the apoptosis of human cervical cancer HeLa cells induced by vitamin K3（VK3）.METHODS： The HeLa cells were divided into 4 groups： control group,30 μmol / L VK3 group,10 mmol / L NH4Cl group and 30 μmol / L VK3 ＋ 10 mmol / L NH4Cl group.The viability of HeLa cells in each group was measured by MTT assay.The changes of lysosomal membrane in HeLa cells were detected by acridine orange（AO） staining.Autophagic vacuoles in the cells were observed by staining with monodansylcadaverine（MDC）.Intracellular acidification in HeLa cells was detected by BCECF-AM flow cytometry analysis.The cells were stained with Hoechst 33342 to determine the chromatin condensation and apoptotic rate.RESULTS： Compared with control group,no effect of NH4Cl on the viability of HeLa cells was observed.In VK3 group,the cell viability decreased.AO staining appeared no change,and the autophagic vacuoles and the cytoplasmic acidification were observed.Nuclear chromatin condensation and cell apoptosis in the cells were also detected.Compared with VK3 group,combination of NH4Cl and VK3 enhanced the lysosomal permeability,the formation of autophagic vacuoles and the cytoplasmic acidification,and exacerbated the apoptosis.CONCLUSION： Autophagolysosomal inhibitor NH4Cl exacerbates apoptosis in HeLa cells,indicating that autophagy exerts protective effect on the process of injury in HeLa cells induced by VK3.

关 键 词：自噬 HELA细胞 氯化铵 维生素K3 

分 类 号：R363.2[医药卫生—病理学]