Enhancement of TAT cell membrane penetration efficiency by dimethyl sulphoxide.
Cell penetrating peptides (CPPs) are promising tools for transducing presynthesized therapeutic molecules which possess low membrane permeability. The poor efficiency of cellular uptake and unexpected cellular localization are still the main obstacles to the development of drug delivery by using CPPs. In this study, we investigated the effect of a penetration enhancer, dimethylsulfoxide (DMSO), on the penetrating efficiency of a synthetic TAT peptide or the TAT fusion protein. FITC-labeled TAT and TAT-GFP were added to 10% DMSO or 100 microM chloroquine pretreated cells, fluorescence uptake into culturing cells was observed using fluorescence microscopy, FACS or quantitatively analyzed by a fluorescence spectrum. 10% DMSO treatment markedly increased internalization of TAT into cells and appeared in a well-distributed pattern throughout the cytosol and nucleus without membrane perforating or detectable cytotoxicity, the enhancement effect by 10% DMSO was reduced by endocytosis inhibitors including ammonium chloride and sodium azide. 10% DMSO also enhanced TAT-Apoptin induced apoptosis of carcinoma cells. These findings implicated that DMSO can be a novel delivery enhancer appropriate for CPP penetration.
Wang
Hu
H
The Institute of Molecular Biology, Three Georges University, Yichang, 443002, P.R. China.
Zhong
Chun-Yan
CY
Wu
Jiang-Feng
JF
Huang
Yu-Bin
YB
Liu
Chang-Bai
CB
D021581:Active Transport, Cell Nucleus; D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002467:Cell Nucleus / Q000378:metabolism; D002469:Cell Separation / Q000379:methods; D002738:Chloroquine / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004705:Endocytosis / Q000187:drug effects; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006461:Hemolysis / Q000187:drug effects; D056945:Hep G2 Cells; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D019810:Sodium Azide / Q000494:pharmacology; D013997:Time Factors; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism*
D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C091717:VP3 protein, Chicken anemia virus; D054322:tat Gene Products, Human Immunodeficiency Virus; D000643:Ammonium Chloride; D049452:Green Fluorescent Proteins; D002738:Chloroquine; D019810:Sodium Azide; D016650:Fluorescein-5-isothiocyanate; D004121:Dimethyl Sulfoxide