摘要
目的观察重组人血管内皮抑素(rhES)联合低氧对人宫颈癌HeLa细胞自噬和凋亡的影响。方法取对数生长期HeLa细胞在低氧的基础上分别给予以下处理:PBS、rhES、rhES联合3-甲基腺嘌呤(3-MA)。处理24 h后,Hochest33528染色及annexin V-FITC/PI染色集合流式细胞术检测细胞的凋亡率,Western blot法检测微管相关蛋白轻链3(LC-3)、Bcl-2、Bax蛋白的表达。结果 Hochest33528染色提示低氧联合rhES组可以诱导细胞出现核碎裂;Annexin V-FITC/PI染色结合流式细胞术检测发现3组细胞的凋亡率分别为:(2.94±0.45)%、(21.38±0.92)%、(6.87±0.58)%,低氧联合rhES诱导细胞凋亡比率明显高于其他2组;Western blot法检测结果显示,与其他2组比较,低氧联合rhES处理HeLa细胞后LC-3Ⅱ及Bax表达水平明显升高,而Bcl-2明显降低。结论 rhES联合低氧可以促使HeLa细胞发生自噬和凋亡。
Objective To observe the impact of recombinant human endostatin（rhES） combined with hypoxia on the autophagy and apoptosis of human cervical cancer HeLa cells. Methods Under hypoxia,HeLa cells in logarithmic growth phase were treated with PBS,rhES,rhES combined with 3-methyladenine（3-MA,inhibitor of autophagy）,respectively.Twenty-four hours later,cell apoptosis rate was detected using Hochest33528 staining and annexin V-FITC/PI staining and flow cytometry; the expressions of microtubule-associated protein light chain 3（LC-3）,Bcl-2,Bax proteins were determined using Western blotting. Results Hochest33528 staining suggested that cells of the hypoxia plus rhES group showed nuclear fragmentation; Annexin V-FITC /PI staining combined with flow cytometry revealed that the apoptosis rates were respectively（2.94 ±0.45）%,（21.38 ±0.92）% and（6.87 ±0.58）% in the three groups. It was significantly higher in the hypoxia plus rhES group than the other two groups. Western blotting showed that the expression levels of LC3（ LC-3 Ⅱ） and Bax increased and Bcl-2 decreased in the hypoxia plus rhES group as compared with the other two groups. Conclusion Endostatin combined hypoxia can induce autophagy and apoptosis of HeLa cells.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2014年第12期1255-1257,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
江苏大学2012年度医学临床科技发展基金(JLY20120158)
关键词
宫颈癌
自噬
重组人血管内皮抑素
低氧
cervical cancer
autophagy
recombinant human endostatin
hypoxia
