中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2010年
16期
1126-1130
,共5页
关翠萍%林福全%洪为松%傅丽芳%周妙妮%许文%许爱娥
關翠萍%林福全%洪為鬆%傅麗芳%週妙妮%許文%許愛娥
관취평%림복전%홍위송%부려방%주묘니%허문%허애아
白癜风%一元酚单氧酶%内质网%FBXO11
白癜風%一元酚單氧酶%內質網%FBXO11
백전풍%일원분단양매%내질망%FBXO11
Vitiligo%Monophenol monooxygenase%Endoplasmic reticulum%FBXO11
目的 通过检测InnVit(即FBX011)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,Lipofectamine TM 2000脂质体方法转染BIOBr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.
目的 通過檢測InnVit(即FBX011)在白癜風患者中的錶達及其對內質網形態和酪氨痠酶輸齣內質網的影響,探討INNVit基因對黑素細胞功能的影響.方法 收集10例錶皮片移植白癜風患者皮損區與正常供皮區組織,免疫組化檢測InnVit蛋白的錶達情況.化學閤成InnVit基因特異性siRNA片段及構建過錶達質粒載體P3XF-P120,Lipofectamine TM 2000脂質體方法轉染BIOBr細胞,電鏡觀察不同處理組細胞的內質網形態變化;激光共聚焦顯微鏡觀察酪氨痠酶蛋白及鈣網蛋白在內質網中的共定位情況;Western印跡檢測轉染細胞中InnVit蛋白、酪氨痠酶及鈣網蛋白的錶達水平.結果 10例白癜風患者皮損區InnVit蛋白錶達明顯低于正常供皮區.未處理組與質粒轉染組的黑素細胞內質網形態正常,而siRNA轉染組的細胞內質網有明顯膨脹,且其相對膨脹率(1.97±0.48)與未處理組(1.28±0.09,P=0.001)和質粒轉染組間(1.24±0.13,P=0.001)比較,差異均有統計學意義.未處理組與質粒轉染組細胞中酪氨痠酶的錶達範圍部分地超齣瞭鈣網蛋白的標記範圍,而siRNA轉染組細胞中酪氨痠酶幾乎與鈣網蛋白共定位于內質網中.siRNA轉染組InnVit蛋白的相對錶達量(0.030±0.004)低于未處理組(0.320±0.020)和質粒轉染組(0.710±0.040),質粒轉染組高于未處理組(均P＜0.01);鈣網蛋白3組間差異無統計學意義(P＞0.05);siRNA轉染組酪氨痠酶(1.040±0.060)高于質粒轉染組(0.720±0.030)和未處理組(0.350±0.030),質粒轉染組高于未處理組(均P＜0.01).結論 白癜風患者皮損區InnVit蛋白錶達下降;InnVit基因的錶達影響瞭黑素細胞內質網的形態,併可以調控酪氨痠酶在內質網中的輸齣,進而影響黑素細胞的功能.
목적 통과검측InnVit(즉FBX011)재백전풍환자중적표체급기대내질망형태화락안산매수출내질망적영향,탐토INNVit기인대흑소세포공능적영향.방법 수집10례표피편이식백전풍환자피손구여정상공피구조직,면역조화검측InnVit단백적표체정황.화학합성InnVit기인특이성siRNA편단급구건과표체질립재체P3XF-P120,Lipofectamine TM 2000지질체방법전염BIOBr세포,전경관찰불동처리조세포적내질망형태변화;격광공취초현미경관찰락안산매단백급개망단백재내질망중적공정위정황;Western인적검측전염세포중InnVit단백、락안산매급개망단백적표체수평.결과 10례백전풍환자피손구InnVit단백표체명현저우정상공피구.미처리조여질립전염조적흑소세포내질망형태정상,이siRNA전염조적세포내질망유명현팽창,차기상대팽창솔(1.97±0.48)여미처리조(1.28±0.09,P=0.001)화질립전염조간(1.24±0.13,P=0.001)비교,차이균유통계학의의.미처리조여질립전염조세포중락안산매적표체범위부분지초출료개망단백적표기범위,이siRNA전염조세포중락안산매궤호여개망단백공정위우내질망중.siRNA전염조InnVit단백적상대표체량(0.030±0.004)저우미처리조(0.320±0.020)화질립전염조(0.710±0.040),질립전염조고우미처리조(균P＜0.01);개망단백3조간차이무통계학의의(P＞0.05);siRNA전염조락안산매(1.040±0.060)고우질립전염조(0.720±0.030)화미처리조(0.350±0.030),질립전염조고우미처리조(균P＜0.01).결론 백전풍환자피손구InnVit단백표체하강;InnVit기인적표체영향료흑소세포내질망적형태,병가이조공락안산매재내질망중적수출,진이영향흑소세포적공능.
Objective To investigate the roles of InnVit(FBX011)gene in melanocytes by detecting the expression of InnVit gene in vitiligo and analyzing the impact of IunVit 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 IunVit gene expression by immunohistochemistry.Synthesimt specific siRNA and constructed plasmid P3XF-P120 were separately transfected into cells for the silence and over-expression of InnVit gene with lipofectamine~(TM) 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.InnVit,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 inilated ER was shown in siRNA group.And the relative inflation rate in siRNA group(1.97±0.48)was higher
