Targeted inhibition of NMYC by peptide nucleic acid in N-myc amplified human neuroblastoma cells: cell-cycle inhibition with induction of neuronal cell differentiation and apoptosis.
We developed an antisense peptide nucleic acid (PNA) targeted against a unique sequence in the terminus of the 5'-UTR of N-myc, designed for selective inhibition of NMYC in neuroblastoma cells. Fluorescent microscopy showed carrier-free delivery of the PNA to two human neuro-blastoma cell lines: GI-LI-N (N-myc-amplified) and GI-CA-N (N-myc-unamplified). Only in the former, PNA treatment determined 70% cell-viability reduction (at 48 h). In N-myc-amplified GI-LI-N cells, the PNA determined NMYC-translation inhibition (Western blotting), accumulation of cells in G1, induction of differentiation and apoptosis. Selectivity of the PNA was demonstrated by altering three point mutations. These findings should encourage development of a PNA-based tumor-specific agent for neuroblastoma (or other neoplasms) with N-myc overexpression.
Pession
Andrea
A
Department of Pediatrics, University of Bologna, S. Orsola Hospital, I-40138 Bologna, Italy. pession@med.unibo.it
Tonelli
Roberto
R
Fronza
Raffaele
R
Sciamanna
Elena
E
Corradini
Roberto
R
Sforza
Stefano
S
Tedeschi
Tullia
T
Marchelli
Rosangela
R
Montanaro
Lorenzo
L
Camerin
Consuelo
C
Franzoni
Monica
M
Paolucci
Guido
G
D017209:Apoptosis; D001483:Base Sequence; D015153:Blotting, Western; D002453:Cell Cycle; D002454:Cell Differentiation; D002455:Cell Division; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000494:pharmacology; D005434:Flow Cytometry; D016193:G1 Phase; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009447:Neuroblastoma / Q000188:drug therapy* / Q000378:metabolism*; D009474:Neurons / Q000473:pathology; D020135:Peptide Nucleic Acids / Q000627:therapeutic use*; D017354:Point Mutation; D016271:Proto-Oncogene Proteins c-myc / Q000037:antagonists & inhibitors* / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D020135:Peptide Nucleic Acids; D016271:Proto-Oncogene Proteins c-myc