The mechanism of polyplex internalization into cells: testing the GM1/caveolin-1 lipid raft mediated endocytosis pathway.
The GM1/caveolin-1 lipid raft mediated endocytosis mechanism was explored for generation 5 and 7 poly(amidoamine) dendrimer polyplexes employing the Cos-7, 293A, C6, HeLa, KB, and HepG2 cell lines. Expression levels of GM1 and caveolin-1 were measured using dot blot and Western blot, respectively. The level of GM1 in the cell plasma membrane was adjusted by incubation with exogenous GM1 or ganglioside inhibitor PPMP, and the level of CAV-1 was adjusted by upregulation with the adenovirus vector expressed caveolin-1 (AdCav-1). Cholera toxin B subunit was employed as a positive control for uptake in all cases. No evidence was found for a GM1/caveolin-1 lipid raft mediated endocytosis mechanism for the generation 5 and 7 poly(amidoamine) dendrimer polyplexes.
Qi
Rong
R
Peking University Institute of Cardiovascular Sciences, Peking University Health Science Center, Peking University, Beijing 100083, China.
Mullen
Douglas G
DG
Baker
James R
JR
Holl
Mark M Banaszak
MM
D000260:Adenoviruses, Human / Q000235:genetics; D000818:Animals; D019556:COS Cells; D051242:Caveolin 1 / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002522:Chlorocebus aethiops; D050091:Dendrimers / Q000008:administration & dosage* / Q000493:pharmacokinetics*; D016503:Drug Delivery Systems; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D005677:G(M1) Ganglioside / Q000378:metabolism*; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D021962:Membrane Microdomains / Q000378:metabolism*; D008614:Meperidine / Q000031:analogs & derivatives / Q000494:pharmacology; D014162:Transfection
C495540:CAV1 protein, human; D051242:Caveolin 1; D050091:Dendrimers; C104700:PAMAM Starburst; C026733:4-propionyloxy-4-phenyl-N-methylpiperidine; D005677:G(M1) Ganglioside; D008614:Meperidine