Hes1在烟草诱导人支气管上皮细胞恶性转化过程中的作用  

Role of Hes1 in malignant transformation of human bronchial epithelial cells induced by tobacco

作　　者：洪磊[1] 周继红[2] 李伟[1] 陈余清[1] 蒋鹏[1] 袁娜娜[1] 王效静[1] 朱茂祥[3] 杨陟华[3] 

机构地区：[1]蚌埠医学院第一附属医院呼吸与危重症医学科,安徽呼吸系病临床基础省级实验室,安徽蚌埠233004 [2]蚌埠医学院生物化学与分子生物学教研室,安徽蚌埠233002 [3]军事医学科学院放射与辐射研究所,北京100850

出　　处：《中国病理生理杂志》2017年第7期1153-1162,共10页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81172213);安徽省自然科学基金资助项目(No.1408085MH14);安徽省高等学校自然科学基金重点项目(No.KJ2016A487);安徽省重点实验室绩效考核补助项目(No.1506c085013);安徽高校自然科学研究重点项目(No.KJ2017A241)

摘　　要：目的:探讨转录因子发状分裂相关增强子1(hairy and enhancer of split,Hes1)在香烟烟气凝集物(cigarette smoke condensate,CSC)诱导永生化人支气管上皮细胞BEP2D恶性转化中的作用。方法:CSC(1L空气中点燃1支香烟)慢性染毒BEP2D细胞至第70代,软琼脂集落形成实验检测CSC诱导的细胞恶性转化表型;采用RT-PCR和Western blot法检测各代细胞的Hes1表达;MTT法、细胞集落形成实验和流式细胞术检测Notch通路阻断剂DAPT或脂质体转染Hes1-siRNA对CSC染毒BEP2D细胞增殖与凋亡的影响。检测吸烟大鼠外周小气道组织中Hes1的表达;采用免疫组化法和RT-PCR法检测非小细胞肺癌组织及正常气道组织中Hes1的表达。结果:第70代BEP2D细胞具备恶性转化表型;Hes1在CSC染毒BEP2D细胞中的表达总体呈逐渐增高的趋势;DAPT和Hes1-siRNA均能通过下调Hes1显著抑制第70代BEP2D细胞的增殖,诱导其凋亡;Hes1在卷烟烟气暴露大鼠气道黏膜1月和6月组的表达较同期对照组显著增高;吸烟显著诱导肺癌组织和正常气道表达Hes1。结论:Hes1可能通过促进凋亡与增殖失衡,参与吸烟诱导的肺癌发生。AIM ： To investigate the role of transcription factor hairy and enhancer of split 1 （ Hesl ） in the ma-lignant transformation of human bronchial epithelial cell line BEP2D induced by tobacco. METHODS： The BEP2D cells were chronically exposed to cigarette smoke condensate （CSC） at 1 cigarette per L until the 70th generation. The pheno-type of malignant transformation of the cells induced by CSC was detected by soft agar clony formation assay. RT-PCR and Western blot were used to determined the expression of Hesl at mRNA and protein levels in each generation of the cells.The proliferation and apoptosis of the BEP2D cells exposed to CSC were analyzed with the methods of MTT assay, flow cy-tometry and cell colony formation assay after treatment with Notch pathway bloker DAPT or liposome transfection with Hesl- siRNA. The expression of Hesl in the peripheral small airway tissues of the smoking rats was evaluated by immunohisto- chemical staining. The expression of Hesl in non-small-cell lung cancer and normal airway tissues was also detected by the methods of immunohistochemistry and RT-PCR. RESULTS： The BEP2D cells in the 70th generation had a malignant transformation phenotype. The expression of Hesl in the BEP2D cells exposed to CSC for different time showed an increasing trend. DAPT and liposome transfection with Hesl-siRNA down-regulated the expression of Hesl, inhibited the cell proliferation and induced cell apoptosis. The expression of Hesl in the airway mucosa of the rats exposed to cigarette smoke for 1 month and 6 months was significantly higher than that in control group. Cigarette smoking induced the expression of Hesl in lung cancer and normal airway tissues. CONCLUSION ： Hesl may be involved in smoking-induced lung cancer by promoting the imbalance between apoptosis and proliferation.

关 键 词：肺癌 吸烟 转录因子Hes1 香烟烟气凝集物 

分 类 号：R730.231[医药卫生—肿瘤] R734.2[医药卫生—临床医学]