發(fā)布時(shí)間：2018-11-25 17:53

【摘要】：目的:探討轉(zhuǎn)錄因子發(fā)狀分裂相關(guān)增強(qiáng)子1(hairy and enhancer of split,Hes1)在香煙煙氣凝集物(cigarette smoke condensate,CSC)誘導(dǎo)永生化人支氣管上皮細(xì)胞BEP2D惡性轉(zhuǎn)化中的作用。方法:CSC(1L空氣中點(diǎn)燃1支香煙)慢性染毒BEP2D細(xì)胞至第70代,軟瓊脂集落形成實(shí)驗(yàn)檢測(cè)CSC誘導(dǎo)的細(xì)胞惡性轉(zhuǎn)化表型;采用RT-PCR和Western blot法檢測(cè)各代細(xì)胞的Hes1表達(dá);MTT法、細(xì)胞集落形成實(shí)驗(yàn)和流式細(xì)胞術(shù)檢測(cè)Notch通路阻斷劑DAPT或脂質(zhì)體轉(zhuǎn)染Hes1-siRNA對(duì)CSC染毒BEP2D細(xì)胞增殖與凋亡的影響。檢測(cè)吸煙大鼠外周小氣道組織中Hes1的表達(dá);采用免疫組化法和RT-PCR法檢測(cè)非小細(xì)胞肺癌組織及正常氣道組織中Hes1的表達(dá)。結(jié)果:第70代BEP2D細(xì)胞具備惡性轉(zhuǎn)化表型;Hes1在CSC染毒BEP2D細(xì)胞中的表達(dá)總體呈逐漸增高的趨勢(shì);DAPT和Hes1-siRNA均能通過(guò)下調(diào)Hes1顯著抑制第70代BEP2D細(xì)胞的增殖,誘導(dǎo)其凋亡;Hes1在卷煙煙氣暴露大鼠氣道黏膜1月和6月組的表達(dá)較同期對(duì)照組顯著增高;吸煙顯著誘導(dǎo)肺癌組織和正常氣道表達(dá)Hes1。結(jié)論:Hes1可能通過(guò)促進(jìn)凋亡與增殖失衡,參與吸煙誘導(dǎo)的肺癌發(fā)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.
【作者單位】： 蚌埠醫(yī)學(xué)院第一附屬醫(yī)院呼吸與危重癥醫(yī)學(xué)科安徽呼吸系病臨床基礎(chǔ)省級(jí)實(shí)驗(yàn)室;蚌埠醫(yī)學(xué)院生物化學(xué)與分子生物學(xué)教研室;軍事醫(yī)學(xué)科學(xué)院放射與輻射研究所;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(No.81172213) 安徽省自然科學(xué)基金資助項(xiàng)目(No.1408085MH14) 安徽省高等學(xué)校自然科學(xué)基金重點(diǎn)項(xiàng)目(No.KJ2016A487) 安徽省重點(diǎn)實(shí)驗(yàn)室績(jī)效考核補(bǔ)助項(xiàng)目(No.1506c085013) 安徽高校自然科學(xué)研究重點(diǎn)項(xiàng)目(No.KJ2017A241)
【分類(lèi)號(hào)】：R734.2

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