[1]朱立强),张 乐),李庆华),等.Smac过表达对食管鳞癌细胞Warburg效应和细胞凋亡的影响[J].郑州大学学报(医学版),2019,(06):789-793.[doi:10.13705/j.issn.1671-6825.2018.11.075]
 ZHU Liqiang),ZHANG Le),LI Qinghua),et al.Effects of Smac overexpression on Warburg effect and apoptosis in esophageal squamous cell carcinoma cells[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2019,(06):789-793.[doi:10.13705/j.issn.1671-6825.2018.11.075]
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Smac过表达对食管鳞癌细胞Warburg效应和细胞凋亡的影响()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2019年06期
页码:
789-793
栏目:
食管癌研究
出版日期:
2019-11-20

文章信息/Info

Title:
Effects of Smac overexpression on Warburg effect and apoptosis in esophageal squamous cell carcinoma cells
作者:
朱立强1)张 乐2)李庆华2)张彦婷2)张璐玉2)刘腾飞2)关方霞2)
1)郑州大学第二附属医院检验科 郑州 450014 2)郑州大学生命科学学院 郑州 450001
Author(s):
ZHU Liqiang1)ZHANG Le2)LI Qinghua2)ZHANG Yanting2) ZHANG Luyu2) LIU Tengfei2) GUAN Fangxia2)
1)Department of Laboratory Medicine, the Second Affiliated Hospital,Zhengzhou University,Zhengzhou 450014 2)College of Life Sciences,Zhengzhou University, Zhengzhou 450001
关键词:
Smac蛋白 食管鳞癌 EC1细胞 Eca109细胞 Warburg效应 细胞凋亡
Keywords:
Smac protein esophageal squamous cell carcinoma EC1 cell Eca109 cell Warburg effect apoptosis
分类号:
R735.1
DOI:
10.13705/j.issn.1671-6825.2018.11.075
摘要:
目的:探究Smac过表达对食管鳞癌细胞Warburg效应和细胞凋亡的影响。方法:以含Smac过表达载体的慢病毒感染食管鳞癌细胞EC1和Eca109(Smac过表达组),以感染含空载体的慢病毒为阴性对照,以未感染细胞为空白对照。采用Western blot法检测3组细胞Smac蛋白的表达情况。用过氧化氢刺激Smac过表达组和阴性对照组细胞,利用线粒体膜电位检测试剂盒和Annexin V-FITC/PI双染法检测细胞线粒体膜电位和细胞凋亡情况,并用Western blot法检测细胞中凋亡信号通路相关蛋白[细胞色素C(Cyt-C)、Bcl-2、Caspase-3前体]以及Warburg效应关键蛋白[己糖激酶Ⅱ(HK2)和乳酸脱氢酶(LDH)]的表达。结果:EC1和Eca109 Smac过表达组Smac蛋白表达水平升高,提示成功构建了Smac过表达细胞株。经过氧化氢刺激后,与阴性对照组相比,Smac过表达组HK2和LDH表达水平下降,细胞线粒体膜电位降低,细胞凋亡率升高,Cyt-C的表达水平升高,Bcl-2和Caspase-3前体蛋白的表达水平下降(P<0.05)。结论:氧化应激刺激下,Smac过表达可抑制食管鳞癌细胞的Warburg效应,促进食管鳞癌细胞凋亡。
Abstract:
Aim:To study the effects of Smac overexpression on Warburg effect and apoptosis in esophageal squamous cell carcinoma(ESCC)cells.Methods:Eca109 and EC1 cells were infected with Smac overexpression lentivirus vector(Smac overexpression group), or mock-vehicle(negative control group), and the untreated cells were blank control group. The expression of Smac was measured by Western blot. The ESCC cells in Smac overexpression group and negative control group were stimulated by hydrogen peroxide,mitochondrial membrane potential was detected and apoptosis was observed by Annexin V-FITC/PI double staining. Finally,Western blot was employed to monitor the expressions of apoptosis signal pathway-related proteins such as cytokines cytochrome C(Cyt-C), Bcl-2,Caspase-3 precursor proteins and Warburg effect-key proteins including hexokinaseⅡ(HK2)and lactate dehydrogenase(LDH).Results:Eca109 and EC1 cells overexpressing Smac were constructed.Compared with negative control group,after hydrogen peroxide stimulation,the expressions of HK2 and LDH reduced in Smac overexpression group, the mitochondrial membrane potential was decreased,additionally, the percentage of apoptotic ESCC cells was significantly enhanced, the expression of Cyt-C was increased, while the expressions of Bcl-2 and Caspase-3 precursor proteins were decreased(P<0.05).Conclusion:Overexpression of Smac would inhibit Warburg effect and promote apoptosis of ESCC cells when treated with oxidative stress.

参考文献/References:

[1] GALLUZZI L,LAROCHETTE N,ZAMZAMI N,et al.Mitochondria as therapeutic targets for cancer chemotherapy[J].Oncogene,2006,25(34):4812
[2] KOPPENOL WH,BOUNDS PL,DANG CV.Otto Warburg's contributions to current concepts of cancer metabolism[J].Nat Rev Cancer,2011,11(5):325
[3] MOHAMED MS,BISHR MK,ALMUTAIRI FM,et al.Inhibitors of apoptosis: clinical implications in cancer[J].Apoptosis,2017,22(12):1487
[4] XU Y,ZHOU L,HUANG J,et al.Role of Smac in determining the chemotherapeutic response of esophageal squamous cell carcinoma[J].Clin Cancer Res,2011,17(16):5412
[5] POFF A,KOUTNIK AP,EGAN KM,et al.Targeting the Warburg effect for cancer treatment: ketogenic diets for check for management of glioma[J].Semin Cancer Biol,2019,56:135
[6] BAYLEY JP,DEVILEE P.The Warburg effect in 2012[J].Curr Opin Oncol,2012,24(1):62
[7] 练雯.优化的反相高效液相色谱法检测能量代谢物质方法的建立和应用[D].广州:广州医学院; 广州医科大学,2009.
[8] BOWMAN CE,ZHAO L,HARTUNG T,et al.Requirement for the mitochondrial pyruvate carrier in mammalian development revealed by a hypomorphic allelic series[J].Mol Cell Biol,2016,36(15):2089
[9] NG H,SMITH DJ,NAGLEY P.Application of flow cytometry to determine differential redistribution of cytochrome c and Smac/DIABLO from mitochondria during cell death signaling[J].PLoS One,2012,7(7):e42298
[10]VERHAGEN AM,EKERT PG,PAKUSCH M,et al.Identification of DIABLO,a mammalian protein that promotes apoptosis by binding to and antagonizing IAP proteins[J].Cell,2000,102(1):43
[11]DU C,FANG M,LI Y,et al.Smac, a mitochondrial protein that promotes cytochrome c-dependent caspase activation by eliminating IAP inhibition[J].Cell,2000,102(1):33
[12]郭彩霞,李艳博,王斌,等.Smac过表达增强氯化镉对人肝癌细胞SMMC-7721的增殖抑制和促凋亡作用[J].吉林大学学报(医学版),2011,37(4):571

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备注/Memo

备注/Memo:
【基金项目】河南省高等学校重点科研项目(15A310028,15A180022); 河南省高校科技创新团队项目(15IRTSTHN022); 河南省科技创新人才计划项目(154200510008) 【作者简介】李庆华,通信作者,女,1983年6月生,博士,讲师,研究方向:肿瘤学,E-mail:guozilqh@126.com; 关方霞,通信作者,女,1969年2月生,博士,教授,研究方向:干细胞与再生医学,E-mail:guanfangxia@126.com
更新日期/Last Update: 2019-11-20