title: Caveolin-1 mediates gene transfer and cytotoxicity of polyethyleneimine in mammalian cell lines.
abstract: Polyethyleneimine (PEI) is a cost-effective and non-viral vector for gene transfer, but the factors determining gene transfer efficiency and cytotoxicity of PEI in different mammalian cell lines remain largely unknown. In the present study, three different cell lines were chosen for investigation. Using pEGFP DNA and PEI, 21.5, 29.2, and 92.1 % of GFP-positive cells were obtained in BMSC, Hela, and 293T, respectively. In luciferase reporter assay, similar results were obtained (for luciferase activity, BMSC < Hela < 293T cells). By MTT test and cell apoptotic marker analysis, we demonstrated that high gene transfer efficiency is accompanied with high cytotoxicity of PEI. Moreover, we found that high expression level of caveolin-1 was accompanied with high gene transfer efficiency and cytotoxicity of PEI in 293T cells. More convincingly, caveolin-1 silencing in 293T could reduce both gene transfer efficiency and cytotoxicity of PEI. In contrast, caveolin-1 overexpression in BMSCs increases both gene transfer efficiency and cytotoxicity of PEI. Taken together, our study suggests that caveolin-1 may at least in part determine gene transfer efficiency and cytotoxicity of PEI in mammalian cell lines, providing caveolin-1 as a potential target for improving gene transfer efficiency when applying positively charged polyplexes to cell transfection.
authors: College of Life Science and Technology, Xinxiang Medical University, 601 Jinsui Road, Xinxiang, 453003, Henan, China.
authors: Hai-Jie
authors: HJ
authors: Yang
authors: Pei
authors: P
authors: Feng
authors: Lei
authors: L
authors: Wang
authors: Zhi-Chao
authors: ZC
authors: Li
authors: Shuang-Ping
authors: SP
authors: Ma
authors: Mian
authors: M
authors: Wang
authors: Zhi-Wei
authors: ZW
authors: Feng
mesh_terms: D000818:Animals; D051242:Caveolin 1 / Q000502:physiology*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D049410:Luciferases, Renilla / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D011094:Polyethyleneimine / Q000633:toxicity*; D014162:Transfection
chemical_list: D051242:Caveolin 1; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine; D049410:Luciferases, Renilla
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