title: Effects of lentiviral vector-mediated TRADD expression on the inhibition of hypertrophic scar formation.
abstract: The tumor necrosis factor receptor-associated death domain protein (TRADD) regulates cell proliferation and apoptosis via tumor necrosis factor alpha (TNF-α)-mediated signaling pathways. Low levels of TRADD expression may result in the excessive proliferation of hypertrophic scar fibroblasts (HSFb). This study investigated the effects of a lentiviral vector carrying the human tradd gene on the proliferation, apoptosis and type I collagen synthesis of HSFb and embryonic fibroblasts (EFb) and further explored the resulting effects on hypertrophic scars (HS). We utilized cytoimmunofluorescence and Western blotting to confirm the expression of TRADD in HSFb and EFb. A PLVX-TRADD-EGFP lentivirus was prepared and transfected into EFb and HSFb, and then the expression of a TRADD-GFP-FLAG fusion protein was detected in HSFb and EFb. After stimulation with 10 ng/mL TNF-α, cell proliferation, apoptosis, and the synthesis of type I collagen were assessed. Our results show that the expression level of TRADD was significantly lower in HSFb than in EFb. A biologically active PLVX-TRADD-EGFP lentivirus was constructed and transfected into HSFb and EFb. The TRADD-GFP-FLAG fusion protein was effectively expressed in HSFb and EFb. Either alone or in combination with 10 ng/mL TNF-α, the PLVX-TRADD-EGFP lentivirus inhibited proliferation, caused a G2/M phase arrest, induced the appearance of a sub-G1 apoptotic peak and inhibited the secretion of type I collagen by HSFb without significantly affecting EFb. These results suggest that the low expression of TRADD in HSFb is a principal reason for their excessive proliferation. The transfection of a PLVX-TRADD-EGFP lentivirus led to the normal expression of TRADD in HSFb. When combined with 10 ng/mL TNF-α, a PLVX-TRADD-EGFP lentivirus transfection could inhibit cell proliferation, promote apoptosis, and reduce the secretion of type I collagen in HSFb, thereby reducing HS formation.
authors: Institute of Gastroenterology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
authors: Yue
authors: Y
authors: Yuan
authors: Institute of Gastroenterology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China pgy63@163.com.
authors: Guiyong
authors: G
authors: Peng
authors: Institute of Gastroenterology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
authors: Xiufeng
authors: X
authors: Kang
authors: Institute of Gastroenterology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
authors: Yunjie
authors: Y
authors: Liu
authors: Institute of Gastroenterology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
authors: Jianhua
authors: J
authors: Dai
authors: Institute of Gastroenterology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
authors: Hongbo
authors: H
authors: Wu
mesh_terms: D017209:Apoptosis; D015153:Blotting, Western; D002460:Cell Line; D049109:Cell Proliferation; D017439:Cicatrix, Hypertrophic / Q000517:prevention & control*; D024042:Collagen Type I / Q000032:analysis; D005347:Fibroblasts / Q000502:physiology*; D020869:Gene Expression Profiling; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008856:Microscopy, Fluorescence; D053199:TNF Receptor-Associated Death Domain Protein / Q000096:biosynthesis* / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D024042:Collagen Type I; D053199:TNF Receptor-Associated Death Domain Protein; D014409:Tumor Necrosis Factor-alpha
keywords: TNF-α; Tumor necrosis factor receptor-associated death domain protein; hypertrophic scars; lentiviral vector