Transfer into a mesothelioma cell line of tumor suppressor gene p16 by cholesterol-based cationic lipids.
In this work, the tumor suppressor gene p16 was efficiently transferred into FR cells isolated from a patient with malignant mesothelioma using cationic liposomes prepared from trimethyl aminoethane carbamoyl cholesterol (TMAEC-Chol) and triethyl aminopropane carbamoyl cholesterol (TEAPC-Chol). This transfer was performed after preliminary assays were undertaken to find the optimal transfection conditions. Results showed that an efficient transfer of plasmids containing the reporter gene pCMV-beta galactosidase vectorized by TMAEC-Chol/DOPE and TEAPC-Chol/DOPE liposomes into mesothelioma FR cells was obtained as assessed by luminometric measurements of beta-galactosidase activity. Cytotoxicity studied by MTT test showed that at concentrations used for this study, the cationic liposomes have no effect on cell growth. Transfer into mesothelioma FR cells of a plasmid construct containing the tumor suppressor gene p16 was carried out with these liposomes. Western blotting and immunofluorescence showed the presence of p16 in treated cells. An inhibition of cell growth was observed, indicating that efficient tumor suppressor gene transfer can be performed by using cationic liposomes.
Piperno-Neumann
S
S
Laboratoire de Spectroscopie Biomoléculaire, CNRS FRE 2313, UFR de Médecine, Université Paris XIII, 74 rue Marcel Cachin, F93017 Bobigny Cedex, France.
Oudar
O
O
Reynier
P
P
Briane
D
D
Cao
A
A
Jaurand
M C
MC
Naejus
R
R
Kraemer
M
M
Breau
J L
JL
Taillandier
E
E
D002455:Cell Division; D002784:Cholesterol / Q000031:analogs & derivatives*; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D018014:Gene Transfer Techniques; D019942:Genes, p16 / Q000502:physiology*; D006801:Humans; D008081:Liposomes; D008654:Mesothelioma / Q000235:genetics* / Q000473:pathology; D010957:Plasmids; D010997:Pleural Neoplasms / Q000235:genetics* / Q000473:pathology; D014162:Transfection; D014407:Tumor Cells, Cultured
C459406:3beta(N-(N',N',N'-triethylaminopropane)carbamoyl)cholesterol iodide; C459405:3beta(N-(N',N',N'-trimethylaminoethane)carbamoyl)cholesterol iodide; D008081:Liposomes; D002784:Cholesterol