A dependence on proteoglycans for cationic lipid-mediated gene transfer has been suggested in previous studies [Mislick and Baldeschwieler (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 12349-12354; Mounkes, Zhong, Cipres-Palacin, Heath and Debs (1998) J. Biol. Chem. 273, 26164-26170]. We have evaluated the mechanism of proteoglycan involvement in cationic lipid-mediated gene transfer. DNA plasmid uptake and gene expression were studied in wild-type Chinese hamster ovary (CHO) cells (CHO-K1), heparan sulphate-deficient CHO cells (pgsD-677) and proteoglycan-deficient CHO cells (pgsB-618). At an optimal ratio of cationic lipid to DNA, a substantial decrease in reporter gene expression was observed in proteoglycan-deficient cells compared with that in heparan sulphate-deficient and wild-type cells. However, there were no differences in reporter gene expression between the cell lines when transfected by electroporation. Moreover, all cell lines exhibited equal cationic-lipid-DNA complex uptake activities, as assessed by the measurement of intracellular 32P-labelled and rhodamine-labelled DNA plasmid. An analysis of reflected-light images of wild-type and proteoglycan-deficient cells suggested that cationic lipids were preferentially toxic to proteoglycan-deficient cells. Cell-growth assays confirmed this, showing that cationic lipids exhibited a greater anti-proliferative activity in proteoglycan-deficient cells and in chlorate-treated wild-type cells than in the other cell lines. The growth-inhibitory effect of cationic lipids was abrogated by the addition of exogenous sulphated glycosaminoglycans. We conclude that the glycosaminoglycan part of proteoglycans serves a protective role against cationic lipid cytotoxicity, allowing optimal transfection efficiency in vitro.
Skip Nav Destination
Article navigation
September 1999
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
Research Article|
August 24 1999
Protective role for proteoglycans against cationic lipid cytotoxicity allowing optimal transfection efficiency in vitro
Mattias BELTING;
Mattias BELTING
1
1Department of Cell and Molecular Biology, Section for Cell and Matrix Biology, Lund University, P.O.B. 94, S-221 00 Lund, Sweden
1To whom correspondence should be addressed (mattias.belting@medkem.lu.se).
Search for other works by this author on:
Per PETERSSON
Per PETERSSON
1Department of Cell and Molecular Biology, Section for Cell and Matrix Biology, Lund University, P.O.B. 94, S-221 00 Lund, Sweden
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
April 29 1999
Revision Received:
June 07 1999
Accepted:
June 30 1999
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1999
1999
Biochem J (1999) 342 (2): 281–286.
Article history
Received:
April 29 1999
Revision Received:
June 07 1999
Accepted:
June 30 1999
Citation
Mattias BELTING, Per PETERSSON; Protective role for proteoglycans against cationic lipid cytotoxicity allowing optimal transfection efficiency in vitro. Biochem J 1 September 1999; 342 (2): 281–286. doi: https://doi.org/10.1042/bj3420281
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Captcha Validation Error. Please try again.