基础医学与临床 ›› 2015, Vol. 35 ›› Issue (6): 792-796.

• 研究论文 • 上一篇    下一篇

过表达Lats1降低食管癌Eca109细胞增殖

罗骏1,王聪懿1,熊伟2,张诚3,吴庆琛3   

  1. 1. 重庆医科大学
    2. 重庆医科大学 附属第一医院 胸心外科
    3. 重庆医科大学附属第一医院
  • 收稿日期:2014-08-29 修回日期:2014-12-01 出版日期:2015-06-05 发布日期:2015-05-27
  • 通讯作者: 吴庆琛 E-mail:qiang196210@163.com

Overexpression of Lats1 reduce proliferation in Eca109 cells

  • Received:2014-08-29 Revised:2014-12-01 Online:2015-06-05 Published:2015-05-27

摘要: 目的 运用慢病毒转染技术在食管癌Eca109细胞中过表达LATS1基因,探究LATS1调控Eca109细胞增殖、凋亡及周期的作用及机制。方法构建LATS1基因的慢病毒载体转染Eca109细胞株,RT-PCR细胞中LATS1mRNA的表达;Western blot检测LATS1、YAP、BAX/BCL-2的蛋白表达水平;MTS检测细胞增殖;流式细胞术检测细胞凋亡及周期;hochest33258观察细胞凋亡染色。结果LATS1慢病毒载体转染Eca109细胞后,目的基因组(Ad-LATS1)LATS1mRNA及LATS1蛋白表达、BAX蛋白表达明显高于对照组(CON)及阴性对照组(Ad-GFP),而YAP、BCL-2的蛋白表达明显低于对照组及Ad-GFP组(P<0.05);Ad-LATS1组Eca109细胞的增殖率从第5天开始明显低于对照组及Ad-GFP组(P<0.05);Ad-LATS1组细胞较Ad-GFP及对照组G1期比例明显增高而S期比例明显缩短,凋亡率明显增高(P<0.05),细胞荧光染色强度及范围也增高。结论LATS1基因可上调BAX、下调YAP、BCL-2使Eca109凋亡,并诱导G1期延长、S期缩短来降低其增殖力。

关键词: [关键字]:食管肿瘤, Lats1, YAP, 增殖 , 凋亡

Abstract: Objective To discuss the impact and mechanism on proliferation,apoptosis and cell cycle in Eca109 cells by overexpress LATS1 in lentivirus transfection technique. Methods Built LATS1-lentivirus vector then transfected the lentivirus vector into Eca109 cells;LATS1 mRNA expression was detected by real-time PCR;LATS1、YAP、BAX/BCL-2 protein expression were detected by western blotting;proliferation ability was detected by MTS assay;the ratio of apoptosis and cell cycle were detected by FCM;the dyeing of apoptosis was observed by hochest33258. Result After transfection of LATS1-lentivirus in Eca109 cells(Ad-LATS1 group),we found lats1 mRNA expression and protein expression level,BAX protein expression level were remarkablely higher than control group and Ad-GFP groups while YAP,BCL-2 protein expression level were on the contrary(P<0.05);The ratio of proliferation in Ad-LATS1 group was notable lower than control group and Ad-GFP groups from the fifth day(P<0.05);FCM showed more cells stay in G1 phase in Ad-LATS1 group than in control group and-GFP groups while the ratio of cells in S phase were on the contrary and apoptosis rate was much higher in Ad-LATS1 group than in others(P<0.05);Hoechst 33258 dying also demonstrated that LATS1 could induce apoptosis in Eca109 cells through the degree and range of dyeing. Conclusion gene LATS1could up-regulate BAX whlle down-regulate YAP and BCL-2 to increase the apoptosis rate of Eca109 cells,more over,it could prolong the G1 phase of cell cycle and shorten the S phase to inhibit the proliferation of Eca109 cells.

Key words: [Key words]:esophagus tumor, Lats1, YAP, proliferation , apoptosis