title: Chondrogenic effect of precartilaginous stem cells following NLS-TAT cell penetrating peptide-assisted transfection of eukaryotic hTGFβ3.
abstract: Cell penetrating peptides (CPPs) are a series of promising carriers for delivering exogenous DNA to living cells. Among them, the combination of the human immunodeficiency virus TAT protein (TAT) with the SV40 large T protein nuclear localization signal (NLS) to form NLS-TAT performs well. In the present study, we took advantage of this new carrier to deliver transforming growth factor-beta 3 (TGFβ3) genes. TGFβ3 was expressed by the pEGFP-N1 vector following transfection of rat precartilaginous stem cells (PSCs), which promoted hTGFβ3 protein self-expression. At 24, 48, 72, and 120 h after transfection, the expression levels of hTGFβ3 were found to be elevated as compared with the control. The expression of hTGFβ3 was found to mediate the chondrogenic effect of PSCs. Thus, we determined the expression of the chondrogenesis-related genes type II collagen, Sox 9, and aggrecan in PSCs at 24, 48, 72, and 120 h after transfection. We found that their transcription and translation was augmented, which indicated a trend of active chondrogenesis in the PSCs. Our results demonstrated that NLS-TAT had the ability to deliver exogenous DNA into rat PSCs and could be actively expressed. This process successfully promoted PSC chondrogenesis. Additionally, PSC, may represent a new type of stem cells, and thus show great potential in regenerative repair following cartilage injury.
authors.affiliation: Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Guo
authors.forename: Xiangyu
authors.initials: X
authors.lastname: Chu
authors.forename: Wenkai
authors.initials: W
authors.lastname: Li
authors.forename: Qiyong
authors.initials: Q
authors.lastname: Pan
authors.forename: Hongbo
authors.initials: H
authors.lastname: You
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000235:genetics / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry; D008856:Microscopy, Fluorescence; D019913:Nuclear Localization Signals / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D014162:Transfection / Q000379:methods*; D053782:Transforming Growth Factor beta3 / Q000235:genetics / Q000378:metabolism*
chemical_list: D019913:Nuclear Localization Signals; D053782:Transforming Growth Factor beta3
keywords: CELL PENETRATING PEPTIDE; CHONDROGENESIS; PEPTIDE-BASED CARRIERS; PRECARTILAGINOUS STEM CELLS