中华皮肤科杂志 ›› 2013, Vol. 46 ›› Issue (12): 858-862.

• 论著 • 上一篇    下一篇

他卡西醇对人表皮黑素细胞增殖、黏附、迁移及c-kit表达的影响

李其林1,何丹华2,牛牧1,黄永华1,陆小娟1   

  1. 1. 暨南大学第四附属医院，广州市红十字会医院皮肤科
    2. 深圳市宝安区妇幼保健院
  • 收稿日期:2012-11-12 修回日期:2013-08-09 发布日期:2013-12-01
  • 通讯作者: 李其林 E-mail:qlli_cn@163.com
  • 基金资助:
    广东省科技计划项目：姜黄素对人表皮黑色素细胞调控作用的研究

Effect of tacalcitol on the proliferation, adhesion, migration and c-kit mRNA expression of human epidermal melanocytes

  • Received:2012-11-12 Revised:2013-08-09 Published:2013-12-01

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摘要： 【摘要】 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-kit mRNA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。 【关键词】 他卡西醇； 黑素细胞； 细胞增殖； 细胞运动； 原癌基因蛋白质c-kit

关键词: 黑素细胞, 他卡西醇, 细胞增殖, 细胞运动, 原癌基因蛋白质c-kit

Abstract: LI Qi-lin, HE Dan-hua, NIU Mu, HUANG Yong-hua, LU Xiao-juan. Department of Dermatology, Fourth Affiliated Hospital of Jinan University/Guangzhou Red Cross Hospital, Guangzhou 510220, China Corresponding author: LI Qi-lin, Email: qlli_cn@163.com 【Abstract】 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 10-8 mol/L for 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 － 10-7 mol/L for 72 hours （all P < 0.01）. Conclusion Tacalcitol can promote melanocytes to proliferate, migrate, express c-kit mRNA, and adhere to fibronectin. 【Key words】 Tacalcitol; Melanocytes; Cell proliferation; Cell movement; Proto-oncogene proteins c-kit

Key words: Tacalcitol

引用本文

李其林 何丹华 牛牧 黄永华 陆小娟. 他卡西醇对人表皮黑素细胞增殖、黏附、迁移及c-kit表达的影响[J]. 中华皮肤科杂志, 2013,46(12):858-862. doi: