Remarkable induction of apoptosis in cancer cells by a novel cationic liposome complexed with a bcl-2 antisense oligonucleotide.
We reported recently a novel cationic cholesterol derivative with a hydroxyethylamino head group, cholesteryl-3beta-carboxyamidoethylene-N-hydroxyethylamine (I) for liposome-mediated gene transfection [FEBS Lett., 408 (1997) 232]. In the present paper we have studied whether this novel cationic liposome is prominent in nature to suppress cell growth of human cancer cells. Bcl-2 antisense phosphorothioate oligonucleotides (AS-ODNs) were complexed with the cationic liposomes with the derivative (I) and they were introduced into human cervix epithelial carcinoma cell lines HeLa, and mouse fibroblast NIH3T3 cells. An AS-ODNs targeting/bcl-2 gene induced probably apoptosis (including necrosis in some cases) in HeLa and NIH3T3 cells, however, nonsense oligonucleotides (NS-ODNs) corresponding to a scrambled-sequence control hardly induced apoptosis. Induction of apoptosis was much greater than that by commercially available DC-Chol liposomes. Fluorescence intensities of FITC-conjugated bcl-2 AS-ODNs were specifically found in the nucleus. The intensity of the AS-ODNs was mostly consistent with the amounts of Bcl-2 proteins observed by Western blot analysis in the target cells. The results showed the possibility that this new cationic cholesterol derivative might be very promising to be used for liposome-mediated gene targeting in vitro and in vivo.
Noguchi
Shoichiro
S
Graduate School of Pharmaceutical Sciences, Nagoya City University, Tanabe-dori, Mizuho-ku, Japan.
Hirashima
Naohide
N
Furuno
Tadahide
T
Nakanishi
Mamoru
M
D017209:Apoptosis / Q000187:drug effects*; D002412:Cations; D004273:DNA, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D005434:Flow Cytometry; D017930:Genes, Reporter / Q000235:genetics; D019254:Genes, bcl-2 / Q000235:genetics*; D006801:Humans; D015151:Immunoblotting; D008081:Liposomes; D008156:Luciferases; D018613:Microscopy, Confocal; D009369:Neoplasms / Q000473:pathology*; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D002412:Cations; D004273:DNA, Neoplasm; D008081:Liposomes; D016376:Oligonucleotides, Antisense; D008156:Luciferases