摘要
目的研究紫外线(UV)照射对皮肤成纤维细胞形态学、生长增殖状态、突变频率和ATPmRNA表达水平的影响。方法采用一定剂量的中长波紫外线照射培养的新生儿包皮成纤维细胞。通过细胞计数记录细胞增殖,光学显微镜观察形态学变化,次黄嘌呤磷酸核糖转移酶突变实验检测突变频率,RT-PCR法检测ATPmRNA表达水平。结果成纤维细胞经UVB照射后,形态受损,细胞增殖减缓,照射72h后细胞活性只增加了约0.2倍,而非照射组增加约2.4倍。在每106个突变克隆细胞中突变频率由20mJ/cm2的(20.4±6.7)个增加到60mJ/cm2的(97.7±7.1)个。ATPmRNA表达水平随照射剂量及照射时间增加而呈下调变化。结论UV照射可使人成纤维细胞ATPmRNA表达水平呈剂量及时间依赖性下调变化,在能量消耗条件下成纤维细胞形态异常,生长受抑,突变频率与照射剂量的增加成正相关。
Objective To investigate the effects of ultraviolet irradiation on the morphology, cell proli-feration, mutation frequency and level of expression of ATP mRNA in human fibroblasts. Methods The infant foreskin fibroblasts were cultured and irradiated with different doses of ultraviolet A or B (UVA or UVB). Light microscopy was used for observing the morphologic change and cell counting of cell proliferation, HPRT-mutagenesis assay for the mutation detection, and RT-PCR for the expression of ATP mRNA. Results Compared with the control group, the irradiated cultured fibroblasts were damaged, cell growth was retarded (MTT assay showed 2.4 fold increase in the control group but only 0.2 fold in the UVB group 72 h after irradiation). Mutation frequency in 106 colony forming cells was increased from 20.4 ± 6.7 at 20 mJ/cm2 to 97.7 ± 7.1 under 60 mJ/cm2 of UVB irradiation. The expression level of ATP mRNA was down-regulated after UV irradiation. There was an increase of mutation frequency which was UVB dose-dependenct, and decrease of ATP mRNA expression which was dose- and time-dependent. Conclusions After UV irradiation of the cultured fibroblasts, the cell damage and growth inhibition are induced, and the mutation frequency is increased in a UVB dose-depen-dent maaner. The expression of ATP mRNA shows down-regulation in both dose- and time-dependent manner.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2004年第7期428-431,共4页
Chinese Journal of Dermatology
基金
国家自然科学基金(30371294)
