Complex formation with plasmid DNA increases the cytotoxicity of cationic liposomes.
Cationic liposomes (CL) are one of the most widely studied non-viral vectors for gene delivery. It is well-known that CL induces cytotoxicity following lipofection. However, little is known regarding the mechanism involved in the cytotoxicity. In this study, the in vitro cytotoxicity of CL and its complex with pDNA (lipoplex) was investigated, and a part of the mechanism of induction as well. While free pDNA did not show any cytotoxicity, pDNA increased the cytotoxicity of CL via the formation of lipoplex. In addition, the lipoplex-induced cytotoxicity increased in a lipoplex dose-dependent manner, irrespective of the type of pDNA, cell line and the absence or presence of serum. An assay showed that apoptosis was largely induced by treatment with the lipoplex (lipofection), but not with CL alone, in the tested range of concentration of CL and pDNA. Furthermore, following treatment with lipoplexes, the cells exhibited the morphological features of apoptosis and DNA fragmentation. A cDNA microarray study showed that the lipofection up-regulated 45 genes related to apoptosis, transcription regulation and immune response. These results clearly indicate that pDNA in the lipoplex increases the cytotoxicity of CL as a result of inducing apoptosis. The fundamental principle for gene therapy is to deliver gene-based therapeutics to target cells for specific gene targeting with minimal cytotoxicity. Our results suggest the possibility that cytotoxicity induced by lipofection, accompanied by gene changes, could intrinsically exacerbate, attenuate or even mask the desired effects of gene-based therapy.
Nguyen
Lap Thi
LT
Department of Pharmacokinetics and Biopharmaceutics, Subdivision of Biopharmaceutical Sciences, Institute of Health Biosciences, The University of Tokushima, Japan.
Atobe
Kazutaka
K
Barichello
Jose Mario
JM
Ishida
Tatsuhiro
T
Kiwada
Hiroshi
H
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002412:Cations / Q000737:chemistry; D045744:Cell Line, Tumor; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D053938:DNA Fragmentation / Q000187:drug effects; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000633:toxicity*; D005910:Glioma / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D008081:Liposomes / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D008546:Melanoma, Experimental / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D010957:Plasmids / Q000378:metabolism*; D051381:Rats; D014162:Transfection
D002412:Cations; D018076:DNA, Complementary; D008081:Liposomes; D004247:DNA