肺癌细胞中SOX1调控ECT2的表达.docVIP

肺癌细胞中SOX1调控ECT2的表达 李宁1 李晓冰2 李素云1 （1河南中医学院第一附属医院，郑州，450008； 2河南中医学院基础医学院，郑州，450008） 摘要 目的 探讨肺癌细胞中SOX1基因是否直接调控ECT2的表达。方法 以肺癌细胞株A549细胞为研究对象，Realtime-PCR检测ECT2表达情况。通过荧光素酶报告检查法以及染色体免疫共沉淀方法检测SOX1基因是否直接调控ECT2的表达。免疫组化检测60例肺癌组织中ECT2与SOX1表达的关系。结果 过表达SOX1后A549细胞中ECT2表达下降，而siRNA抑制SOX1后ECT2表达升高，荧光素酶报告检查法以及染色体免疫共沉淀方法提示SOX1可直接与ECT2启动子结合，抑制ECT2的表达。SOX1与ECT2在肺癌组织中表达负相关(r=-0.433，P =0.001)。结论 肺癌细胞A549中SOX1基因直接调控ECT2的表达。 关键词 肺癌；SOX1；ECT2 SOX1 is a novel transcriptional repressor for ECT2 in human lung cancer Li Ning1 Li Xiao-bing2 Li Su-yun1 The First Affiliated Hospital of Henan University of Traditional Chinese Medicine;2 Basic Medical College of Henan University of Traditional Chinese Medicine, China ABSTRACT Objective To test whether SOX1 is a novel transcriptional repressor for ECT2 in human lung cancer. Methods The expression of ECT2 in the A549 cell was detected by Realtime-PCR. We determined whether SOX1 may regulate the expression of ECT2 by using Chromin immunoprecipitation (ChIP) assay and Luciferase activity assay. Immunohistochemical analysis was used to test whether there is an inverse correlation between ECT2 and SOX1. Results Ectopic expression of SOX1 in the A549 cell repressed ECT2 expression. Conversely, siRNA silencing of the SOX1 gene increased ECT2 expression. We also show that SOX1 directly interacted with and repressed the ECT2 promoter. Moreover, the analysis of 60 primary lung cancer samples revealed an inverse correlation between ECT2 and SOX1 levels(r=-0.433，P =0.001). Conclusions SOX1 is a novel transcriptional repressor for ECT2 in human lung cancer. [KEY WORDS] Lung cancer; SOX1; ECT2 肺癌是严重危害人类生命和健康的恶性肿瘤之一，也是世界范围内发病和死亡率最高的癌症。以顺铂为基础的化疗方案是目前针对NSCLC主要的治疗方法，然而，肺癌细胞对顺铂产生的耐药是困扰临床的难题[1]。SOX1，即SRY (sex determining region Y)-box 1，属于具有high mobility group (HMG)-box结构的转录因子家族SOX家族的成员。我们前期结果发现SOX1基因在A549细胞中处于非甲基化状态，在耐顺铂细胞株A549/cis细胞中处于高甲基化状态。抑制SOX1表达可通过激活肺癌细胞自噬能力，从而导致肺癌细胞耐药[2]。前期实验我们通过cDNA微阵列分析发现过表达SOX1后ECT2基因表达降低，本研究我们将探讨肺癌细胞中SOX1基因是否直接调控ECT2的表达。