目的观察甲醛在低剂量水平时短期作用对细胞增殖及DNA损伤的影响,为进一步开展甲醛致体外细胞恶性转变的长期试验提供剂量设计依据。方法采用CCK-8法检测细胞活力,采用BrdU掺入法检测S期细胞比例,从而反映细胞增殖潜能,应用单细胞凝胶电泳结合紫外线照射的方法检测细胞DNA损伤程度。结果在24 h的作用时间下,当甲醛浓度在10-9~10-5mol/L范围内时可以增加细胞增殖率,提高16HBE细胞活力,而剂量达到10-4mol/L时可造成细胞死亡增多,降低细胞活力;同时当甲醛浓度超过10-6mol/L时可产生DNA损伤,在10-6~10-5mol/L表现为DNA断裂,当剂量达到或高于10-4mol/L,DNA蛋白质交联严重;甲醛在10-7~10-5mol/L浓度范围内可以显著增加S期细胞含量,从而促进16HBE细胞增殖。结论 10-6~10-5mol/L甲醛可以在造成16HBE细胞损伤的同时,干扰细胞周期、促进细胞增殖。
Objective To observe the short term effect on cell proliferation and DNA damage of low dose formaldehyde(FA),and provide some fundamental data for the design of in vitro malignant transformation.Methods The method of CCK-8 was applied to detect the cell viability,and BrdU incorporation method was used to observe the percentage of S-phase cells.The degree of DNA damage was determined by the combination of the test of single cell gel electrophoresis and UV exposure.Results On the background of 24h exposure time,when FA concentration ranged from 10-9 to 10-5mol/L it could increase the proliferation ratio and cell viability of 16HBE cells,and cell viability decreased while the concentration reached to 10-4mol/L;DNA damage was observed in the groups greater than 10-6mol/L,while DNA strand break happened in the 10-6~10-5mol/L groups and DNA protein cross-link happened in the groups higher than 10-4mol/L.The percentage of S-phase cell was marked increased on the influence of 10-7~10-5mol/L FA,and then the cell proliferation was promoted.Conclusions The FA could disturb the cell cycle,and then induce the cell proliferation and cause DNA damage simultaneously in 16HBE cells when its concentration ranged within 10-6~10-5mol/L.