Regulation of Mcl-1 by constitutive activation of NF-kappaB contributes to cell viability in human esophageal squamous cell carcinoma cells

BMC Cancer

Bangliang Yin¹  Jianguo Hu¹  Ya Cao³  Yifeng Yang¹  Xinmin Zhou¹  Fenglei Yu¹  Qin Wu¹  Sufeng Ouyang¹  Jian Wang²  Xiaolong Ma¹  Li Xie¹  Mingjiu Chen¹  Zhenkun Xia¹  Yunchang Yuan¹  Jinfu Yang²  Haidan Liu² 

[1]Department of Cardiothoracic Surgery, The Second Xiangya Hospital, Central South University, 139 Renmin Road, Changsha, Hunan 410011, China

[2]Clinical Center for Gene Diagnosis and Therapy, The Second Xiangya Hospital, Central South University, 139 Renmin Road, Changsha, Hunan 410011, China

[3]Cancer Research Institute, Xiangya School of Medicine, Central South University, 110 Xiangya Road, Changsha, Hunan 410078, China

关键词: Cell viability;    Mcl-1;    NF-κB;    Gene regulation;    Esophageal squamous cell carcinoma;   

Others  :  859059
DOI  :  10.1186/1471-2407-14-98

 received in 2013-05-17, accepted in 2014-02-11,  发布年份 2014

【 摘 要 】

Background

Esophageal squamous cell carcinoma (ESCC) is one of the most lethal malignancies with a 5-year survival rate less than 15%. Understanding of the molecular mechanisms involved in the pathogenesis of ESCC becomes critical to develop more effective treatments.

Methods

Mcl-1 expression was measured by reverse transcription (RT)-PCR and Western blotting. Human Mcl-1 promoter activity was evaluated by reporter gene assay. The interactions between DNA and transcription factors were confirmed by electrophoretic mobility shift assay (EMSA) in vitro and by chromatin immunoprecipitation (ChIP) assay in cells.

Results

Four human ESCC cell lines, TE-1, Eca109, KYSE150 and KYSE510, are revealed increased levels of Mcl-1 mRNA and protein compare with HaCaT, an immortal non-tumorigenic cell line. Results of reporter gene assays demonstrate that human Mcl-1 promoter activity is decreased by mutation of kappaB binding site, specific NF-kappaB inhibitor Bay11-7082 or dominant inhibitory molecule DNMIkappaBalpha in TE-1 and KYSE150 cell lines. Mcl-1 protein level is also attenuated by Bay11-7082 treatment or co-transfection of DNMIkappaBalpha in TE-1 and KYSE150 cells. EMSA results indicate that NF-kappaB subunits p50 and p65 bind to human Mcl-1-kappaB probe in vitro. ChIP assay further confirm p50 and p65 directly bind to human Mcl-1 promoter in intact cells, by which regulates Mcl-1 expression and contributes to the viability of TE-1 cells.

Conclusions

Our data provided evidence that one of the mechanisms of Mcl-1 expression in human ESCC is regulated by the activation of NF-kappaB signaling. The newly identified mechanism might provide a scientific basis for developing effective approaches to treatment human ESCC.

【 授权许可】

   
2014 Liu et al.; licensee BioMed Central Ltd.

【 图 表 】

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