Pession Andrea, Tonelli Roberto, Fronza Raffaele, Sciamanna Elena, Corradini Roberto, Sforza Stefano, Tedeschi Tullia, Marchelli Rosangela, Montanaro Lorenzo, Camerin Consuelo, Franzoni Monica, Paolucci Guido
Department of Pediatrics, University of Bologna, S. Orsola Hospital, I-40138 Bologna, Italy.
Int J Oncol. 2004 Feb;24(2):265-72.
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.
我们研发了一种反义肽核酸（PNA），其靶向作用于N-myc 5'-非翻译区（UTR）末端的独特序列，旨在选择性抑制神经母细胞瘤细胞中的NMYC。荧光显微镜显示，PNA可在无载体的情况下递送至两种人类神经母细胞瘤细胞系：GI-LI-N（N-myc扩增）和GI-CA-N（N-myc未扩增）。仅在前一种细胞系中，PNA处理导致细胞活力降低70%（48小时时）。在N-myc扩增的GI-LI-N细胞中，PNA可抑制NMYC翻译（蛋白质免疫印迹法），使细胞在G1期积累，诱导分化和凋亡。通过改变三个点突变证明了PNA的选择性。这些发现应会促进开发一种基于PNA的、针对N-myc过表达的神经母细胞瘤（或其他肿瘤）的肿瘤特异性药物。
