Regulation of Mcl-1 by constitutive activation of NF-κB contributes to cell viability in human esophageal squamous cell carcinoma cells.
Regulation of Mcl-1 by constitutive activation of NF-κB contributes to cell viability in human esophageal squamous cell carcinoma cells.
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DOI：
10.1186/1471-2407-14-98
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发表时间：
2014-02-17
期刊：
影响因子：
3.8
通讯作者：
Yin B
中科院分区：
文献类型：
--
作者：
Liu H;Yang J;Yuan Y;Xia Z;Chen M;Xie L;Ma X;Wang J;Ouyang S;Wu Q;Yu F;Zhou X;Yang Y;Cao Y;Hu J;Yin B
食管鳞状细胞癌（ESCC）是恶性肿瘤中最致命的一种，其5年生存率低于15%。了解ESCC发病机制的分子机制对于开发更有效的治疗方法至关重要。Mcl-1的表达通过逆转录（RT）-PCR和Western blotting测定。报告基因检测法检测人Mcl-1启动子活性。DNA和转录因子之间的相互作用通过体外电泳迁移率变动分析（EMSA）和细胞染色质免疫沉淀（ChIP）分析来证实。4种人食管鳞癌细胞系TE-1、Eca 109、KYSE 150和KYSE 510的Mcl-1 mRNA和蛋白表达水平均高于HaCaT（一种永生的非致瘤细胞系）。报告基因分析的结果表明，在TE-1和KYSE 150细胞系中，人Mcl-1启动子活性通过kappaB结合位点、特异性NF-κ B抑制剂Bay 11 -7082或显性抑制分子DNMI kappaB α的突变而降低。Mcl-1蛋白水平也通过Bay 11 -7082处理或在TE-1和KYSE 150细胞中共转染DNMI κ B α而减弱。EMSA结果表明，NF-κ B亚基p50和p65在体外与人Mcl-1-κ B探针结合。ChIP实验进一步证实了p50和p65在完整细胞中直接与人Mcl-1启动子结合，通过其调节Mcl-1的表达并有助于TE-1细胞的存活。我们的数据提供的证据表明，Mcl-1的表达在人类食管鳞癌的机制之一是通过激活NF-κ B信号调节。新发现的机制可能为开发治疗人类ESCC的有效方法提供科学依据。
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. 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. 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. 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.
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作者：
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