Zeng Wei, Li Hui, Chen Yan, Lv Hongbo, Liu Ling, Ran Jihua, Sun Xiaohong, Bieerkehazhi Shayahati, Liu Yining, Li Xiaomiao, Lai Wenting, Watibieke Jibieke, Dawulietihan Meiliwuerti, Li Xiumei, Li Huiwu
Department of Labour Hygiene and Sanitary Science, College of Public Health, Xinjiang Medical University, Ürümqi, Xinjiang Uyghur Autonomous Region 830011, P.R. China.
Department of Biochemistry and Molecular Biology, School of Basic Medicine, Xinjiang Medical University, Ürümqi, Xinjiang Uyghur Autonomous Region 830011, P.R. China.
Mol Med Rep. 2016 Feb;13(2):1869-80. doi: 10.3892/mmr.2015.4737. Epub 2015 Dec 30.
Survivin and transcription factor p65 (NF‑κB p65) participate in the progression of esophageal squamous cell carcinoma (ESCC). However, the mechanism of NF‑κB p65 activation in ESCC remains to be elucidated. The aim of the present study was to investigate the role of survivin in the activation of NF‑κB p65 in ESCC. The expression levels of survivin, NF‑κB p65, inhibitor of nuclear factor κB kinase subunit α (IKKα) and inhibitor of nuclear factor κB kinase subunit β (IKKβ) were detected in ESCC tissue samples. Eca109 and KYSE150 cells were cultured and survivin activity was modulated via transfection with an overexpression plasmid, a small hairpin RNA plasmid and a specific inhibitor. Quantitative reverse transcription-polymerase chain reaction and western blotting assays were conducted to assess the effects of survivin on the expression levels of IKKα, IKKβ and NF‑κB p65. Cell cycle and apoptosis assays were conducted to detect surviving-dependent cellular behavior changes. In addition, the luciferase reporter gene assay and chromatin immunoprecipitation assay were conducted to determine the genomic sites responsible for surviving-induced activation of NF‑κB p65. The present study demonstrated that the expression of survivin is positively correlated with IKKα and IKKβ in ESCC tissues. Survivin affected the mRNA and protein expression levels of IKKα, IKKβ, and NF‑κB p65 in Eca109 and KYSE150 cells. Furthermore, survivin increased the transcriptional activity of the IKKβ promoter and bound to the IKKβ promoter region in the Eca109 cells. Downregulation of survivin arrested the cell cycle at the G2/M phase and induced apoptosis. Results of the present study suggest that survivin activates NF‑κB p65 in Eca109 cells via binding to the IKKβ promoter region and upregulating IKKβ promoter transcriptional activity. Survivin overexpression activates NF‑κB p65, which is important in the acquisition and maintenance of the oncogenic characteristics of ESCC.
生存素与转录因子p65（核因子κB p65，NF-κB p65）参与食管鳞状细胞癌（ESCC）的进展。然而，ESCC中NF-κB p65激活的机制仍有待阐明。本研究的目的是探讨生存素在ESCC中NF-κB p65激活中的作用。检测ESCC组织样本中生存素、NF-κB p65、核因子κB激酶亚基α（IKKα）抑制剂和核因子κB激酶亚基β（IKKβ）抑制剂的表达水平。培养Eca109和KYSE150细胞，并通过转染过表达质粒、小发夹RNA质粒和特异性抑制剂来调节生存素活性。进行定量逆转录-聚合酶链反应和蛋白质印迹分析，以评估生存素对IKKα、IKKβ和NF-κB p65表达水平的影响。进行细胞周期和凋亡分析，以检测生存素依赖性细胞行为变化。此外，进行荧光素酶报告基因分析和染色质免疫沉淀分析，以确定负责生存素诱导的NF-κB p65激活的基因组位点。本研究表明，ESCC组织中生存素的表达与IKKα和IKKβ呈正相关。生存素影响Eca109和KYSE150细胞中IKKα、IKKβ和NF-κB p65的mRNA和蛋白质表达水平。此外，生存素增加了IKKβ启动子的转录活性，并在Eca109细胞中与IKKβ启动子区域结合。生存素的下调使细胞周期停滞在G2/M期并诱导凋亡。本研究结果表明，生存素通过与IKKβ启动子区域结合并上调IKKβ启动子转录活性来激活Eca109细胞中的NF-κB p65。生存素的过表达激活NF-κB p65，这在ESCC致癌特征的获得和维持中很重要。
