Effect of vinblastine on transfection: influence of cell types, cationic lipids and promoters.
As previously shown, vinblastine, when incorporated into a cationic lipid prior to generation of lipoplexes, increases by approximately 30-fold the extent of transfection of pbeta-Gal with a cytomegalovirus promoter (pCMV-beta-Gal) to vascular smooth muscle cells (VSMC) by 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine (EDOPC)-pCMV-beta-Gal complexes. To test if this increase is limited to VSMC and EDOPC, or is general, we examined three other cell types, human umbilical artery endothelial cells (HUAEC), baby hamster kidney (BHK) cells and 293 cells derived from human kidney, as well as a different cationic lipid, 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP). In addition, to determine the contribution of the NF-kappaB transcription factor to the vinblastine effect, pCMV was replaced with a smooth muscle gamma-actin gene promoter, SMGA, which, unlike pCMV, does not respond to NF-kappaB. It was found that on all cell types we tested, the transfection efficiency increased with vinblastine incorporation; however, the magnitude depended greatly on the cell type, e.g. whereas the transfection of VSMC increased approximately 30-fold, that of 293 cells increased only approximately 2-fold. The cationic phospholipid could be replaced with DOTAP with no loss of effect. In contrast, the promoter was critical and the stimulation was lost if pCMV was replaced with pSMGA. It is concluded that the positive effect of vinblastine on transfection is general and the stimulation of the transcription factor NF-kappaB is involved in this action. The activation of NF-kappaB by anti-microtubule agents should thus allow for transfection of specific cell types by vinblastine lipoplexes.
Wang
Li
L
Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, IL 60208, USA. l-wang4@northwestern.edu
Dean
David A
DA
Macdonald
Robert C
RC
D000199:Actins / Q000235:genetics; D000818:Animals; D002412:Cations / Q000737:chemistry; D002460:Cell Line; D002478:Cells, Cultured; D006224:Cricetinae; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008055:Lipids / Q000737:chemistry; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection / Q000379:methods*; D014747:Vinblastine / Q000493:pharmacokinetics / Q000494:pharmacology*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D000199:Actins; D002412:Cations; D005229:Fatty Acids, Monounsaturated; D008055:Lipids; D016328:NF-kappa B; D000644:Quaternary Ammonium Compounds; D011993:Recombinant Fusion Proteins; D014747:Vinblastine; D001616:beta-Galactosidase; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane