CAJ | 학술논문

目的探讨他卡西醇对体外培养的人表皮黑素细胞增殖、黏附、迁移及c-kit mRNA相对表达的影响.方法用不同浓度的他卡西醇干预体外培养的人表皮黑素细胞,采用四唑氮氢氧化物(XTT)法分别检测培养24、48、72 h后黑素细胞的增殖活性;用纤维连接蛋白(FN)包被细胞培养板检测培养72 h后黑素细胞的黏附率;用Transwell微孔膜法检测培养24 h后黑素细胞在FN上的迁移情况;逆转录-聚合酶链反应(RT-PCR)检测培养72 h后黑素细胞c-kit mRNA相对表达量.采用重复测量方差分析及完全随机设计方差分析进行统计学检验.结果重复测量方差分析结果显示,10-10、10-9、10-8、10-7、10-6 mol/L他卡西醇促进黑素细胞增殖作用的差异有统计学意义(F=9.47,P＜ 0.01),上述浓度他卡西醇在培养24、48、72 h后促进黑素细胞增殖作用的差异有统计学意义(F=14.44,P＜ 0.01),药物浓度和培养时间之间存在交互作用(F=2.47,P＜0.01),其中10-8 mol/L他卡西醇与黑素细胞共同培养72 h黑素细胞增殖活性最高.10-8～10-7 mol/L他卡西醇可显著促进黑素细胞在FN上的黏附(均P＜ 0.01);10-9 ～ 10-8 mol/L他卡西醇在培养24 h能显著促进黑素细胞迁移(均P＜ 0.01);RT-PCR显示10-9 ～ 10-7 mol/L他卡西醇在培养72 h均能显著增加黑素细胞c-kitmRNA的相对表达量(均P＜ 0.01).结论他卡西醇可促进培养的人表皮黑素细胞增殖及在FN上的黏附、迁移作用,并可上调黑素细胞c-kit mRNA的表达.
목적 탐토타잡서순대체외배양적인표피흑소세포증식、점부、천이급c-kit mRNA상대표체적영향.방법 용불동농도적타잡서순간예체외배양적인표피흑소세포,채용사서담경양화물(XTT)법분별검측배양24、48、72 h후흑소세포적증식활성;용섬유련접단백(FN)포피세포배양판검측배양72 h후흑소세포적점부솔;용Transwell미공막법검측배양24 h후흑소세포재FN상적천이정황;역전록-취합매련반응(RT-PCR)검측배양72 h후흑소세포c-kit mRNA상대표체량.채용중복측량방차분석급완전수궤설계방차분석진행통계학검험.결과 중복측량방차분석결과현시,10-10、10-9、10-8、10-7、10-6 mol/L타잡서순촉진흑소세포증식작용적차이유통계학의의(F=9.47,P＜ 0.01),상술농도타잡서순재배양24、48、72 h후촉진흑소세포증식작용적차이유통계학의의(F=14.44,P＜ 0.01),약물농도화배양시간지간존재교호작용(F=2.47,P＜0.01),기중10-8 mol/L타잡서순여흑소세포공동배양72 h흑소세포증식활성최고.10-8～10-7 mol/L타잡서순가현저촉진흑소세포재FN상적점부(균P＜ 0.01);10-9 ～ 10-8 mol/L타잡서순재배양24 h능현저촉진흑소세포천이(균P＜ 0.01);RT-PCR현시10-9 ～ 10-7 mol/L타잡서순재배양72 h균능현저증가흑소세포c-kitmRNA적상대표체량(균P＜ 0.01).결론 타잡서순가촉진배양적인표피흑소세포증식급재FN상적점부、천이작용,병가상조흑소세포c-kit mRNA적표체.
Objective To evaluate the effect of tacalcitol on the proliferation,adhesion,migration and c-kit mRNA expression of cultured human epidermal melanocytes.Methods Cultured epidermal melanocytes from the prepuce of adolescent males were treated with various concentrations of tacalcitol.Then,cell proliferation was evaluated by tetrazolium salt (XTT) assay after 24,48 and 72 hours of treatment,adhesive activity by using fibronectin-coated culture plates after 72 hours,migratory activity by Transwell assay using a microporous membrane after 24 hours,and the c-kit mRNA expression was semiquantitatively analyzed by reverse transcription PCR after 72 hours of treatment.Statistical analysis was done by repeated-measure analysis of variance and completely random design analysis of variance.Results As repeated-measure analysis of variance showed,tacalcitol of 10-10,10-9,10-8,10-7 and 10-6 mol/L significantly promoted the proliferation of melanocytes (F =9.47,P ＜ 0.01),with significant differences in the promoting effect among various durations of treatment with different concentrations of tacalcitol (F =14.44,P ＜ 0.01),and with significant interaction effect between drug concentration and treatment duration (F =2.47,P ＜ 0.01).The highest proliferation level was observed in melanocytes treated with tacalcitol of 10-s mol/Lfor 72 hours.There was a significant increase in the adhesion rate of human epidermal melanocytes to fibronectin after treatment with tacalcitol of 10-8-10-7 mol/L for 72 hours (both P ＜ 0.01),number of melanocytes migrating through micropore membranes per high-power field (× 200) after treatment with tacalcitol of 10-9-10-8 mol/L for 24 hours (both P ＜ 0.01),and in the c-kit mRNA expression in melanocytes treated with tacalcitol of 10-9-10-7mol/L for 72 hours (all P ＜ 0.01).Conclusion Tacalcitol can promote melanocytes to proliferate,migrate,express c-kit mRNA,and adhere to fibronectin.