MS-275对食管鳞癌KYSE-70细胞存活、细胞周期、凋亡及迁移的影响  

Effects of MS-275 on survival,cell cycle,apoptosis and migration of human esophageal squamous cancer KYSE-70 cells

作　　者：刘腾飞 周建康 黄团结 王亚苹 周馨魁 张乐 张璐玉 陈一豪 刘红涛 关方霞 马珊珊 LIU Tengfei;ZHOU Jiankang;HUANG Tuanjie;WANG Yaping;ZHOU Xinkui;ZHANG Le;ZHANG Luyu;CHEN Yihao;LIU Hongtao;GUAN Fangxia;MA Shanshan(College of Life Sciences,Zhengzhou University,Zhengzhou 450001)

机构地区：[1]郑州大学生命科学学院,郑州450001

出　　处：《郑州大学学报（医学版）》2018年第5期547-552,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金资助项目(81601078;81471306);河南省高校科技创新团队支持计划(15IRTSTHN022);河南省科技创新人才计划(154200510008)

摘　　要：目的:观察MS-275对食管鳞癌KYSE-70细胞存活、细胞周期、凋亡以及迁移的影响,并分析其对PI3K/Akt/m TOR信号通路相关蛋白p-Akt1和p-m TOR的影响。方法:采用qRT-PCR和Western blot检测KYSE-70细胞和正常食管上皮Het-1A细胞中HDAC1 mRNA及蛋白的表达;CCK-8法检测不同浓度(0.25、0.50、1.00、2.00、4. 00、8. 00μmol/L) MS-275对KYSE-70细胞存活的影响;以0. 00、0. 25、0. 50、1. 00和2. 00μmol/L MS-275处理KYSE-70细胞,48 h后流式细胞仪检测细胞周期,Annexin V/PI染色检测细胞凋亡,划痕实验检测细胞迁移情况,Western blot检测细胞中Cyclin D1、Cleaved caspase-3、E-cadherin、p-Akt1和p-mTOR蛋白的表达情况。结果:与Het-1A细胞相比,KYSE-70细胞中HDAC1 mRNA和蛋白表达显著增加(P <0. 05);MS-275对KYSE-70细胞存活的影响具有时间和剂量依赖性(P <0. 05);随着MS-275处理浓度的增加,KYSE-70 G0/G1期细胞增加、S期细胞降低,细胞凋亡率提高,划痕愈合率降低(P <0. 05)。MS-275可提高Cleaved caspase-3和E-cadherin蛋白的表达,降低Cyclin D1、p-Akt1和p-mTOR蛋白的表达(P <0. 05)。结论:MS-275可降低KYSE-70细胞存活率,有效抑制细胞迁移,阻滞细胞于G0/G1期,促进细胞凋亡,其作用可能与PI3K/Akt/mTOR信号通路的抑制有关。Aim：To investigate the effects of MS-275 on the cell survival,apoptosis,cell cycle and migration of human esophageal squamous cancer KYSE-70 cells and analyze PI3 K/Akt/m TOR signaling pathway related protein expression.Methods： The expression levels of HDAC1 mRNA and protein in KYSE-70 cells and normal esophageal epithelial cells（Het-1 A） were respectively detected by qRT-PCR and Western blot; the survival of KYSE-70 cells was detected by CCK-8 to determine the optimal time and concentration;KYSE-70 cells were treated with 0. 00,0. 25,0. 50,1. 00,2. 00 μmol/L MS-275 for 48 h,then the apoptosis and the change of cell cycle were detected by flow cytometry; cell migration ability was detected by cell scratches experiment; Cyclin D1,Cleaved caspase-3,E-cadherin and the key proteins（ p-Akt1 and pm TOR） of PI3 K/Akt/m TOR signaling pathway were detected by Western blot. Results：HDAC1 mRNA and protein were overexpressed in KYSE-70 cells（ P〈0. 05） compared with Het-1 A cells;CCK-8 assay showed that the survival of KYSE-70 cells were inhibited by MS-275 in a dose-and time-dependent manner（ P〈0. 05）;with the increase of the concentration of MS-275,the apoptosis rate of KYSE-70 cells was significantly increased,the proportion of cells in G0/G1 phase was increased,that of S phase cells was decreased,and the number of migrating cells was significantly decreased（ P〈0. 05）;the relative contents of Cleaved caspase-3 and E-cadherin protein were increased,but those of Cyclin D1,p-Akt1 and p-m TOR protein were reduced by MS-275（ P〈0. 05）. Conclusion：MS-275 can effectively inhibit survival and migration,induce apoptosis,arrest cycle of KYSE-70 cells,which might be related with the inhibition of PI3 K/Akt/m TOR signaling pathway.

关 键 词：MS-275 食管鳞癌 组蛋白去乙酰化酶 PI3K/Akt/m TOR 

分 类 号：R735.1[医药卫生—肿瘤]