title: Construction of human LRIG1-TAT fusions and TAT-mediated LRIG1 protein delivery.
abstract: Human leucine-rich repeats and immunoglobulin-like domains (LRIG1) is a tumor suppressor in animals and also functions as an endogenous suppressor in human tumor. The level of LRIG1 expression is highly associated with patient survival in clinic. The exploration of LRIG1 as a protein drug is an important task. HIV-1 transactivator of transcription peptide (TAT) is an excellent candidate for protein transduction. In this study, human LRIG1 was cloned and LRIG1-TAT fusion gene was constructed. The fusion proteins were produced by an Escherichia coli strain and purified by Ni(2+)-resin. Western blot assay and immunofluorescence microscopy were employed for monitoring LRIG1-TAT protein transduction into human neuroblastoma cells. Cell proliferation and invasion were measured for evaluating the effect of LRIG1-TAT on neuroblastoma cell. Our data showed that LRIG1 protein can be delivered into cells or organs in living animals by TAT. One-time transduction of LRIG1 proteins into human neuroblastoma cells enhanced cell proliferation and increased cell invasion. In vivo transduction showed that LRIG1-TAT protein can be presented in living animal organs. Our experiments provide a new vision on LRIG1 applications and also offer a therapy window for revealing the intrinsic function of LRIG1 on cells.
authors: Department of Neurosurgery, The Oilfield General Hospital of Daqing, Daqing 163001, China.
authors: Yuchun
authors: Y
authors: Wang
authors: Department of Neurosurgery, The Oilfield General Hospital of Daqing, Daqing 163001, China.
authors: Liqi
authors: L
authors: Fu
authors: Department of Neurosurgery, The Oilfield General Hospital of Daqing, Daqing 163001, China.
authors: Bo
authors: B
authors: Liu
authors: Department of Neurosurgery, The Oilfield General Hospital of Daqing, Daqing 163001, China.
authors: Xiaomin
authors: X
authors: Wang
authors: Department of Neurosurgery, The Oilfield General Hospital of Daqing, Daqing 163001, China.
authors: Kai
authors: K
authors: Wang
authors: The First affiliated Hospital of Soochow University, Suzhou 215006, China. Electronic address: yeming@suda.edu.cn.
authors: Ming
authors: M
authors: Ye
mesh_terms: D000818:Animals; D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D016503:Drug Delivery Systems; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D006801:Humans; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000494:pharmacology*; D009361:Neoplasm Invasiveness; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D014161:Transduction, Genetic; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000494:pharmacology*
chemical_list: C484445:LRIG1 protein, human; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus
keywords: Cell invasion; HIV-1 transactivator of transcription peptide; Human leucine-rich repeats and immunoglobulin-like domains; Protein transduction; TAT-mediated protein delivery