308nm单频准分子光对人黑素细胞凋亡、周期和KIT蛋白表达的影响  

Effect of monochromatic 308 nm excimer light on ceil cycle, apoptosis of and KTT expression in human melanocytes

作　　者：张勇[1] 项蕾红[1] 李剑[1] 祝绿川[1] 郑志忠[1] 

机构地区：[1]复旦大学附属华山医院皮肤科,上海200040

出　　处：《中华皮肤科杂志》2008年第3期160-162,共3页Chinese Journal of Dermatology

基　　金：国家自然科学基金（30471560）

摘　　要：目的 探讨308nm单频准分子光对体外培养的人黑素细胞凋亡、细胞周期和KIT蛋白表达的影响。方法 在流式细胞仪上采用AnnexinV检测0，20，40，80，160，300mJ／cm^2剂量308nm单频准分子光照射后24h黑素细胞的凋亡；采用碘化丙啶检测80mJ／cm^2照射后24h细胞周期的变化。实时荧光PCR和Werstern印迹分别检测0，20，40，80，160，300mJ／cm^2剂量照射后24h黑素细胞c—kit mRNA和KIT蛋白的表达。结果照射后24h，在小于160mJ／cm。剂量下，黑素细胞凋亡无明显增加，但300mJ／cm^2时的凋亡率与对照相比较明显增加（P〈0．05）。80mJ／cm^2剂量照射后24h，S期细胞数明显高于其对照，分别为29．06％±0．75％和22．64％±0．07％；低剂量308nm单频准分子光照射后24h，c—kitmRNA表达呈先增加后减少的趋势，80mJ／cm^2时达高峰，KIT蛋白的表达与mRNA的表达趋势一致。结论低剂量308nm单频准分子光（80mJ／cm^2）促进黑素细胞增殖和KIT的表达。Objective To evaluate the effects of 308 nm monochromatic excimer light （MEL） on cell apoptosis, cycle of and KIT expression in human melanocytes. Methods Melanocytes were obtained from circumcised prepuce, and subjected to primary culture. After 4-5 passages, the cells were irradiated with 308 nm MEL at the doses of 0, 20, 40, 80, 160 and 300 mJ/cm^2. After another 24-hour culture, flow cytometric analysis was used to detect the apoptosis and life cycle of these cells, real time RT-PCR and Westemblot to measure the expressions of c-kit mRNA and KIT protein in melanocytes, respectively. Results The apoptosis of melanocytes was not significantly changed after exposure to MEL at doses lower than 160 mJ/cm^2, but was increased significantly by the irradiation of MEL at the dose of 300 mJ/cm^2 （P 〈 0.05 ）. After exposure to 308 nm MEL at the dose of 80 mJ/cm^2, the percentage of melanocytes in S phase were （29.06 ± 0.75）%, significantly higher than that in the untreated group （22.64 ± 0.07）% （P 〈 0.05）. The expression of c-kit mRNA was significantly increased by MEL at the doses of 40 and 80 mJ/cm^2, but decreased significantly at the doses of 160 and 300 mJ/cm^2. The changes of KIT protein were consistent with those of c-kit mRNA after exposure to 308 nm MEL. Conclusion The 308 nm MEL at the dose of 80 mJ/cm^2 can promote the proliferation of and expression of KIT protein in melanocytes, which may be a mechanism for the treatment ofvitiligo with 308 nm MEL.

关 键 词：准分子光 黑素细胞 细胞凋亡 细胞周期 原癌基因蛋白质c—kit 

分 类 号：R758.41[医药卫生—皮肤病学与性病学]