title: Biophysical and biochemical characterization of a liposarcoma-derived recombinant MnSOD protein acting as an anticancer agent.
abstract: A recombinant MnSOD (rMnSOD) synthesized by specific cDNA clones derived from a liposarcoma cell line was shown to have the same sequence as the wild-type MnSOD expressed in the human myeloid leukaemia cell line U937, except for the presence of the leader peptide at the N-terminus. These results were fully confirmed by the molecular mass of rMnSOD as evaluated by ES/MS analysis (26662.7 Da) and the nucleotide sequence of the MnSOD cDNA. The role of the leader peptide in rMnSOD was investigated using a fluorescent and/or (68)Gallium-labelled synthetic peptide. The labelled peptide permeated MCF-7 cells and uptake could be inhibited in the presence of an excess of oestrogen. In vivo it was taken up by the tumour, suggesting that the molecule can be used for both therapy and diagnosis. The in vitro and in vivo pharmacology tests confirmed that rMnSOD is only oncotoxic for tumour cells expressing oestrogen receptors. Pharmacokinetic studies in animals performed with (125)I- and (131)I-labelled proteins confirmed that, when administered systemically, rMnSOD selectively reached the tumour, where its presence was unambiguously demonstrated by scintigraphic and PET scans. PCR analysis revealed that Bax gene expression was increased and the Bcl2 gene was down regulated in MCF7 cells treated with rMnSOD, which suggests that the protein induces a pro-apoptotic mechanism.
authors.affiliation: Department of Molecular Biology and Biotherapy, National Cancer Institute G. Pascale Naples, Naples, Italy. aldo_mancini@tiscali.it
authors.forename: Aldo
authors.initials: A
authors.lastname: Mancini
authors.forename: Antonella
authors.initials: A
authors.lastname: Borrelli
authors.forename: Antonella
authors.initials: A
authors.lastname: Schiattarella
authors.forename: Luigi
authors.initials: L
authors.lastname: Aloj
authors.forename: Michela
authors.initials: M
authors.lastname: Aurilio
authors.forename: Franco
authors.initials: F
authors.lastname: Morelli
authors.forename: Alessandra
authors.initials: A
authors.lastname: Pica
authors.forename: Antonella
authors.initials: A
authors.lastname: Occhiello
authors.forename: Roberto
authors.initials: R
authors.lastname: Lorizio
authors.forename: Roberto
authors.initials: R
authors.lastname: Mancini
authors.forename: Alessandro
authors.initials: A
authors.lastname: Sica
authors.forename: Lelio
authors.initials: L
authors.lastname: Mazzarella
authors.forename: Filomena
authors.initials: F
authors.lastname: Sica
authors.forename: Paolo
authors.initials: P
authors.lastname: Grieco
authors.forename: Ettore
authors.initials: E
authors.lastname: Novellino
authors.forename: Daniela
authors.initials: D
authors.lastname: Pagnozzi
authors.forename: Piero
authors.initials: P
authors.lastname: Pucci
authors.forename: Jean
authors.initials: J
authors.lastname: Rommelaere
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology* / Q000627:therapeutic use*; D001483:Base Sequence; D001669:Biochemical Phenomena; D001671:Biochemistry; D055592:Biophysical Phenomena; D001703:Biophysics; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity; D002942:Circular Dichroism; D004958:Estradiol / Q000494:pharmacology; D006262:Health; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D008080:Liposarcoma / Q000188:drug therapy / Q000201:enzymology* / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D017421:Sequence Analysis; D021241:Spectrometry, Mass, Electrospray Ionization; D013482:Superoxide Dismutase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D023041:Xenograft Model Antitumor Assays
chemical_list: D000970:Antineoplastic Agents; D011994:Recombinant Proteins; D004958:Estradiol; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase