Hes1在烟草诱导人支气管上皮细胞恶性转化过程中的作用

发布时间：2018-11-25 17:53

【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。
[Abstract]:Aim: to investigate the role of transcription factor hair-division related enhancer 1 (hairy and enhancer of split,Hes1) in the malignant transformation of immortalized human bronchial epithelial cells (BEP2D) induced by cigarette smoke agglutination (cigarette smoke condensate,CSC). Methods: CSC (1L (1 cigarette was lit in the air) chronic exposure to BEP2D cells to the 70th generation, soft Agar colony formation assay to detect the CSC induced cell malignant transformation phenotype, RT-PCR and Western blot methods were used to detect the expression of Hes1. MTT assay, colony forming assay and flow cytometry were used to detect the effects of Notch pathway blocker DAPT or liposome transfection of Hes1-siRNA on the proliferation and apoptosis of BEP2D cells exposed to CSC. The expression of Hes1 in peripheral small airway tissues of smoking rats and the expression of Hes1 in non-small cell lung cancer tissues and normal airway tissues were detected by immunohistochemical method and RT-PCR method. Results: the BEP2D cells of passage 70 had malignant transformation phenotype, the expression of Hes1 in BEP2D cells exposed to CSC was gradually increasing, DAPT and Hes1-siRNA could significantly inhibit the proliferation and induce apoptosis of BEP2D cells through down-regulation of Hes1. The expression of Hes1 in the airway mucosa of rats exposed to cigarette smoke in 1 month and 6 months was significantly higher than that in the control group, and smoking significantly induced the expression of Hes1. in lung cancer tissues and normal airway. Conclusion: Hes1 may be involved in the development of lung cancer induced by smoking by promoting the imbalance of apoptosis and proliferation.
【作者单位】：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;蚌埠医学院生物化学与分子生物学教研室;军事医学科学院放射与辐射研究所;
【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)
【分类号】：R734.2

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