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目的通过香烟烟雾体外染毒诱发细胞恶性转化,建立吸烟致肺癌的细胞模型。方法确定合适染烟浓度及时间后,将永生化人支气管上皮细胞系BEAS-2B细胞分为对照组(C)和20%烟雾浓度组(S)。以每孔1×105细胞接种于Transwell培养皿,贴壁后置于气体染毒装置中,香烟烟雾持续泵入直接对细胞染毒。检测染毒后5、10、15、20代细胞恶性转化情况,流式细胞仪分析细胞周期和凋亡变化。结果 BEAS-2B细胞在20%烟雾浓度下染毒5代后,出现生长动力学和形态学改变,生长失去接触抑制,获得对血清诱导分化的抗性和锚着独立性生长能力。G1期细胞百分比明显降低,S期细胞百分比明显升高,G2期细胞百分比先升高后降低,各代细胞凋亡率均明显降低。结论香烟烟雾染毒能够诱发BEAS-2B细胞体外转化,且随染毒代数的增加呈现剂量效应关系,可作为体外研究吸烟致肺癌的细胞模型。
Objective To induce cell malignant transformation by cigarette smoke exposure in vitro and establish a cell model of lung cancer caused by smoking. METHODS: BEAS-2B cells were divided into control group (C) and 20% smoke concentration group (S). Inoculated 1 × 105 cells per well in Transwell culture dishes, adherent placed in a gas-exposed device, cigarette smoke continued to pump directly to the cells. The malignant transformation of cells was detected on day 5, 10, 15 and 20 after infection. The cell cycle and apoptosis were analyzed by flow cytometry. Results After BEAS-2B cells were exposed to 20% smoke for 5 generations, growth kinetics and morphological changes were observed in BEAS-2B cells. Exposure to BEAS-2B cells lost contact inhibition and acquired resistance to serum-induced differentiation and anchorage-independent growth. The percentage of cells in G1 phase was significantly decreased, the percentage of cells in S phase was significantly increased, the percentage of cells in G2 phase was increased first and then decreased, and the apoptosis rate in each passage was significantly decreased. Conclusions Cigarette smoke exposure can induce BEAS-2B cells to transform in vitro and show a dose-response relationship with increasing exposure to algebra. Therefore, BEAS-2B cells can be used as a cell model for the study of smoking-induced lung cancer in vitro.