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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 the transcription factor hairy and enhancer of split 1 (Hes1) in the malignant transformation of immortalized human bronchial epithelial cells (BEP2D) induced by cigarette smoke condensate (CSC). Methods: CSC (1 cigarette lit in the air) chronic exposure to BEP2D cells to the 70th generation, soft agar colony formation assay was used to detect CSC-induced malignant transformation phenotype; using RT-PCR and Western blot detection of each passage cells Hes1 expression; MTT assay, colony formation assay and flow cytometry were used to detect Notch pathway blocker DAPT or liposome-transfected Hes1-siRNA on CSC-infected BEP2D cells proliferation and apoptosis. The expression of Hes1 was detected in small airway tissues of smoking rats. The expression of Hes1 in non-small cell lung cancer tissues and normal airway tissues was detected by immunohistochemistry and RT-PCR. Results: The 70th generation BEP2D cells had a malignant transformation phenotype. The expression of Hes1 in CSE-infected BEP2D cells tended to increase gradually. Both DAPT and Hes1-siRNA could significantly inhibit the proliferation of BEP2D cells on the 70th generation by down- Induced Hes1 expression in airway mucosa of rats exposed to cigarette smoke for 1 month and 6 months respectively, which was significantly higher than that of the control group. Smoking induced significant Hes1 expression in lung cancer tissues and normal airway. Conclusion: Hes1 may be involved in cigarette smoking-induced lung cancer by promoting imbalance between apoptosis and proliferation.