silencing of gpnmb by sirna inhibits the formation of melanosomes in melanocytes in a mitf-independent fashion沉默的gpnmb核抑制黑色素细胞中黑素体的形成一个mitf-independent时尚.pdfVIP

Silencing of GPNMB by siRNA Inhibits the Formation of Melanosomes in Melanocytes in a MITF-Independent Fashion 1,2. 1,2 . 3 1 1 1 1 Ping Zhang , Wei Liu * , Cansheng Zhu , Xiaoying Yuan , Dongguang Li , Weijie Gu , Huimin Ma , Xin Xie4, Tianwen Gao2 1 Department of Dermatology, the General Hospital of the Air Force, Beijing, China, 2 Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi’an, China, 3 Shaanxi Provincial Institute for Endemic Disease Control, Xi’an, China, 4 Reg Lab of Tissue Engineering, Faculty of Life Science, Northwest University, Xi’an, China Abstract Background: Melanosomes are specialized membrane-surrounded organelles, which are involved in the synthesis, storage and transport of melanin. Glycoprotein (transmembrane) non-metastatic melanoma protein b (GPNMB), a melanosome- specific structural protein, shares significant amino acid sequence homology with Pmel-17. Proteomic analysis demonstrated that GPNMB is present in all stages (I-IV) of melanosomes. However, little is known about the role of GPNMB in melanosomes. Methodology/Principal Findings: Using real-time quantitative PCR, Western blotting and immunofluorescence analysis, we demonstrated that the expression of GPNMB in PIG1 melanocytes was up-regulated by ultraviolet B (UVB) radiation. Transmission electron microscopy analysis showed that the total number of melanosomes in PIG1 melanocytes was sharply reduced by GPNMB-siRNA transfection. Simultaneously, the expression levels of tyrosinase (Tyr), tyrosinase related protein 1 (Trp1), Pmel17/gp100 and ocular albinism type 1 protein (OA1) were all significantly attenuated. But the express