摘要
目的观察转谷氨酰胺酶1基因(transglutaminase 1,TGM1)表达沉默前、后对角质形成细胞的分化及角质化包膜形成相关蛋白表达的影响。方法应用免疫细胞化学SP法和Western印迹法检测RNA干扰(RNAi)技术沉默TGM1基因表达前、后角质形成细胞分化标志物K10、内披蛋白、丝聚蛋白及角质化包膜形成相关的兜甲蛋白和富含脯氨酸的小蛋白(small proline-rich proteins,SPRRs)的表达差异。结果免疫细胞化学检测显示:TGM1沉默后细胞K10、兜甲蛋白和富含脯氨酸的小蛋白呈弱阳性表达或阴性表达,而内披蛋白和丝聚蛋白呈中等阳性表达或强阳性表达;Western印迹法检测结果显示,TGM1沉默后K10、兜甲蛋白和富含脯氨酸的小蛋白的蛋白条带亮度明显降低,而内披蛋白和丝聚蛋白条带亮度与空白对照组和阴性对照组相比没有明显改变。结论 TGM1基因沉默可能通过抑制K10的表达影响表皮细胞分化;通过抑制角质化包膜形成相关蛋白兜甲蛋白和富含脯氨酸的小蛋白的表达影响表皮细胞角质化包膜的形成。
Objective To explore the effects of the transglutaminase 1 （ TGM1 ） gene silencing by small interfering RNA （siRNA） in immortalized human keratinocyte（ HaCaT cells） on cell differentiation and expression of comifled envelope related proteins. Methods Before and after RNA interference （RNAi） silencing TGM1 gene, cell differentiation and expression of cornified envelope related proteins were detected by immunocytochemistry and Western blotting. Results Inmmnohistochemical results indicated that cell differentiation and comifled envelope related proteins K10, Loricrin and small proline-rich proteins （ SPRRs ） were negative after transfecting with TGM1 siRNA. Western blotting indicated that very low endogenous expression of the K10. Loricrin and small proline-rich proteins （SPRRs） proteins was observed after transfected with TGM1 siRNA. Conclusion It suggests that TGM1 expression in HaCaT cells are closely related with the cell differentiation and cornified envelope.
出处
《中国皮肤性病学杂志》
CAS
CSCD
北大核心
2016年第2期123-127,共5页
The Chinese Journal of Dermatovenereology
基金
国家自然科学基金面上项目(81071286)
广东省自然科学基金(10151009503000002)
广州市科技计划项目(12C33151651)
广州市医药卫生科技重点项目(201102A212016)
