InnVit／FBXO11在白癜风中的表达及其对酪氨酸酶输出内质网的影响  

Expression of InnVit/FBXOll in vitiligo and its role in tyrosinase export from endoplasmic reticulum

作　　者：关翠萍[1] 林福全[2] 洪为松[1] 傅丽芳[1] 周妙妮[1] 许文[1] 许爱娥[1] 

机构地区：[1]安徽医科大学杭州市第三人民医院皮肤科,杭州310009 [2]浙江理工大学生物工程研究所

出　　处：《中华医学杂志》2010年第16期1126-1130,共5页National Medical Journal of China

基　　金：国家自然科学基金（30671896、30872280）;浙江省自然科学基金（Z206607）

摘　　要：目的通过检测InnVit（即FBXO11）在白癜风患者中的表达及其对内质网形态和酪氨酸酶输出内质网的影响，探讨InnVit基因对黑素细胞功能的影响。方法收集10例表皮片移植白癜风患者皮损区与正常供皮区组织，免疫组化检测InnVit蛋白的表达情况。化学合成InnVit基因特异性siRNA片段及构建过表达质粒载体P3XF-P120，npofectamine^TM 2000脂质体方法转染B10Br细胞，电镜观察不同处理组细胞的内质网形态变化；激光共聚焦显微镜观察酪氨酸酶蛋白及钙网蛋白在内质网中的共定位情况；Western印迹检测转染细胞中InnVit蛋白、酪氨酸酶及钙网蛋白的表达水平。结果10例白癜风患者皮损区InnVit蛋白表达明显低于正常供皮区。未处理组与质粒转染组的黑素细胞内质网形态正常，而siRNA转染组的细胞内质网有明显膨胀，且其相对膨胀率（1．97±0．48）与未处理组（1．28±0．09，P=0．001）和质粒转染组间（1．24±0．13，P=0．001）比较，差异均有统计学意义。未处理组与质粒转染组细胞中酪氨酸酶的表达范围部分地超出了钙网蛋白的标记范围，而siRNA转染组细胞中酪氨酸酶几乎与钙网蛋白共定位于内质网中。siRNA转染组InnVit蛋白的相对表达量（0．030±0．004）低于未处理组（0．320±0．020）和质粒转染组（0．710±0．040），质粒转染组高于未处理组（均P〈0．01）；钙网蛋白3组间差异无统计学意义（P〉0．05）；siRNA转染组酪氨酸酶（1．040±0．060）高于质粒转染组（0．720±0．030）和未处理组（0．350±0．030），质粒转染组高于未处理组（均P〈0．01）。结论白癜风患者皮损区InnVit蛋白表达下降；InnVit基因的表达影响了黑素细胞内质网的形态，并可以调控酪氨酸酶在内质网中的输出，进而影响黑素细胞的功能。Objective To investigate the roles of IunVit （FBXO11） gene in melanocytes by detecting the expression of InnVit gene in vitiligo and analyzing the impact of InnVit gene on morphology of endoplasmic reticulum （ER） and the tyrosinase export from ER. Methods The lesion tissues and the donor tissues were collected from 10 vitiligo patients to examine the InnVit gene expression by immunohistochemistry. Synthesized specific siRNA and constructed plasmid P3XF-P120 were separately transfected into cells for the silence and over-expression of InnVit gene with lipofectamineTM 2000. The untreated cells were used as control. Morphology of ER of cells from the above three groups was observed under electron microscope. Co-localization of tyrosinase and calreticulin was identified by confocal laser scanning microscopy. IunVit, tyrosinase and calreticulin were examined by Western blot. Results In vitiligo patients, the expression of InnVit gene in the lesions was markedly lower than that in the donor tissues. The normal morphology of ER was found in the untreated and the plasmid groups whereas inflated ER was shown in siRNA group. And the relative inflation rate in siRNA group （ 1.97 ± 0. 48 ） was higher than that in the untreated group（ 1.28 ± 0. 09） and plasmid group （1.24 ± 0. 13 ） （both P = 0. 001 ）. In the untreated and the plasmid groups, tyrosinase was expressed beyond the scope marked by ER marker protein calretieulin partly, but co-localized with ealreticulin in ER in the siRNA group. Western blot showed that, contrast to the untreated group （ 0. 320 ± 0. 020 ）, a lower expression level of InnVit in the siRNA group （0. 030 ± 0. 004, P = 0. 001 ） and a higher expression of InnVit in the plasmid group were shown （ 0. 710 ± 0. 040,P =0. 001 ）. No significant difference about the expression level of calretieulin was observed among the three groups （P 〉 0. 05）. As compared with the untreated group（ 0. 350 ±0. 030）, a higher tyrosinase level in the siRNA group （ 1. 040

关 键 词：白癜风 一元酚单氧酶 内质网 FBXO11 

分 类 号：R758.41[医药卫生—皮肤病学与性病学]