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目的探讨在亚砷酸盐致人支气管上皮(HBE)细胞恶性转化过程中氧化应激指标的变化趋势,初步探讨其对细胞恶性转化的作用。方法用1μmol/L亚砷酸钠连续染毒HBE细胞。通过形态学观察、基质金属蛋白酶-9(MMP-9)以及软琼脂克隆实验鉴定细胞恶性转化情况,分别于暴露1、8、15、22、29、36、43代,以流式细胞仪检测细胞活性氧(ROS)水平,检测细胞丙二醛(MDA)含量和超氧化物歧化酶(SOD)活力。结果亚砷酸钠长期染毒HBE细胞至43代时,细胞生长速度明显加快,且呈浸润型生长,MMP-9分泌量升高,细胞集落数量增多。与正常对照(0代)比较,22代及以前HBE细胞内ROS、MDA及SOD的水平均呈先升高后下降的波浪形趋势;29代及以后,ROS、MDA水平逐渐下降,且以ROS表现更为明显,而SOD水平呈逐渐升高趋势。结论细胞内氧化-抗氧化平衡失调可能在低剂量砷化合物诱导HBE细胞恶性转化过程中发挥重要作用。
Objective To investigate the change trend of oxidative stress in the process of malignant transformation of arsenite-induced human bronchial epithelial cells (HBE), and to explore its effect on malignant transformation of cells. Methods HBE cells were exposed continuously with 1 μmol / L sodium arsenite. Malignant transformation was identified by morphological observation, matrix metalloproteinase-9 (MMP-9) and soft agar cloning experiments. The cells were exposed to 1, 8, 15, 22, 29, 36, and 43 passages for detection by flow cytometry The level of reactive oxygen species (ROS), the content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) were measured. Results When sodium arsenite was exposed to HBO for 43 passages, the growth rate of HBE cells was significantly increased. The growth rate of MMP-9 increased and the number of colonies increased. Compared with the normal control (0 generation), the levels of ROS, MDA and SOD in HBE cells at 22nd generation and before were both increased and then decreased. At 29th generation and later, the levels of ROS and MDA gradually decreased, The performance is more obvious, while the level of SOD showed a gradual upward trend. Conclusion The imbalance of intracellular oxidation-antioxidant balance may play an important role in the malignant transformation of HBE cells induced by low doses of arsenic compounds.