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目的 从煤烟对细胞膜理化特性的影响 ,对DNA的损伤 ,诱导原癌基因表达方面研究其致癌作用机制。方法 利用 1,6 二苯基 1,3,5 己三烯 (DPH)和N 3芘 马来酰亚胺 (N 3P M)分别标记NIH3T3细胞膜脂和膜蛋白 ,不同剂量的煤烟作用后 ,通过其荧光偏振度的改变确定膜理化特性的变化 ;利用溴化乙锭 (EB)作荧光探针 ,通过DNA EB结合物荧光强度的变化确定基因组DNA损伤程度 ;利用地高辛标记的c myc癌基因作探针 ,运用斑点杂交的方法 ,检测c myc基因的表达情况。结果 不同剂量的煤烟作用后 ,能够引起膜脂流动性升高 ,膜蛋白运动度无显著变化 ,能够引起DNA的断裂损伤及c myc基因的高表达 ,且上述变化皆与剂量密切相关。结论 煤烟的诱变性与其对细胞膜、DNA及原癌基因的表达的影响密切相关。
Objective To study the mechanism of carcinogenesis by studying the effects of soot on the physicochemical properties of cell membrane, DNA damage, and induction of proto-oncogene expression. METHODS: Membrane lipids and membrane proteins of NIH3T3 cells were labeled with 1,6-diphenyl 1,3,5-hexatriene (DPH) and N 3 芘-maleimide (N 3P M), respectively. After different doses of soot, Changes in the physicochemical properties of the membrane were determined by changes in the degree of polarization of the fluorescence; ethidium bromide (EB) was used as a fluorescent probe to determine the degree of genomic DNA damage by changes in the fluorescence intensity of DNA EB conjugates; the digoxin labeled c myc was used. The oncogene was used as a probe and dot-blot hybridization was used to detect the expression of c myc gene. Results After different doses of soot, the membrane lipid fluidity was increased, the membrane protein mobility was not changed significantly, and DNA fragmentation damage and c myc gene expression could be induced. These changes were closely related to the dose. Conclusion The mutagenicity of soot is closely related to its influence on the expression of cell membrane, DNA and proto-oncogene.