title: Theranostic properties of a survivin-directed molecular beacon in human melanoma cells.
abstract: Survivin is an inhibitor of apoptosis overexpressed in different types of tumors and undetectable in most terminally differentiated normal tissues. In the current study, we sought to evaluate the in vitro theranostic properties of a molecular beacon-oligodeoxynucleotide (MB) that targets survivin mRNA. We used laser scanning confocal microscopy to study MB delivery in living cells and real-time PCR and western blot to assess selective survivin-targeting in human malignant melanoma cells. We further assess the pro-apoptotic effect of MB by measuring internucleosomal DNA fragmentation, dissipation of mitochondrial membrane potential (MMP) and changes in nuclear morphology. Transfection of MB into A375 and 501 Mel cells generated high signal intensity from the cytoplasm, while no signal was detected in the extracellular environment and in survivin-negative cells (i.e., human melanocytes and monocytes). MB time dependently decreased survivin mRNA and protein expression in melanoma cells with the maximum effect reached at 72 h. Treatment of melanoma cells with MB induced apoptosis by significant changes in MMP, accumulation of histone-complexed DNA fragments in the cytoplasm and nuclear condensation. MB also enhanced the pro-apoptotic effect of standard chemotherapeutic drugs tested at clinically relevant concentrations. The MB tested in the current study conjugates the ability of imaging with the pharmacological silencing activity against survivin mRNA in human melanoma cells and may represent an innovative approach for cancer diagnosis and treatment.
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Carpi
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Fogli
authors.affiliation: Institute of Applied Physics "Nello Carrara," IFAC-CNR, Sesto Fiorentino, Florence, Italy.
authors.forename: Ambra
authors.initials: A
authors.lastname: Giannetti
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Adinolfi
authors.affiliation: Institute of Applied Physics "Nello Carrara," IFAC-CNR, Sesto Fiorentino, Florence, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Tombelli
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Da Pozzo
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Alessia
authors.initials: A
authors.lastname: Vanni
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Enrica
authors.initials: E
authors.lastname: Martinotti
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Martini
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Maria Cristina
authors.initials: MC
authors.lastname: Breschi
authors.affiliation: Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa, Pisa, Italy.
authors.forename: Mario
authors.initials: M
authors.lastname: Pellegrino
authors.affiliation: Department of Pharmacy, University of Pisa, Pisa, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Nieri
authors.affiliation: Institute of Applied Physics "Nello Carrara," IFAC-CNR, Sesto Fiorentino, Florence, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Baldini
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D053938:DNA Fragmentation / Q000187:drug effects; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000096:biosynthesis / Q000235:genetics*; D008545:Melanoma / Q000473:pathology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects / Q000235:genetics; D015345:Oligonucleotide Probes / Q000235:genetics / Q000378:metabolism* / Q000627:therapeutic use; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D000077022:Survivin
chemical_list: D000970:Antineoplastic Agents; C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D000077022:Survivin