N-Myc knockdown and apigenin treatment controlled growth of malignant neuroblastoma cells having N-Myc amplification

Gene. 2013 Oct 15;529(1):27-36. doi: 10.1016/j.gene.2013.07.094. Epub 2013 Aug 11.

Abstract

Malignant neuroblastomas mostly occur in children and are frequently associated with N-Myc amplification. Oncogene amplification, which is selective increase in copy number of the oncogene, provides survival advantages in solid tumors including malignant neuroblastoma. We have decreased expression of N-Myc oncogene using short hairpin RNA (shRNA) plasmid to increase anti-tumor efficacy of the isoflavonoid apigenin (APG) in human malignant neuroblastoma SK-N-DZ and SK-N-BE2 cell lines that harbor N-Myc amplification. N-Myc knockdown induced morphological and biochemical features of neuronal differentiation. Combination of N-Myc knockdown and APG most effectively induced morphological and biochemical features of apoptotic death. This combination therapy also prevented cell migration and decreased N-Myc driven survival, angiogenic, and invasive factors. Collectively, N-Myc knockdown and APG treatment is a promising strategy for controlling the growth of human malignant neuroblastoma cell lines that harbor N-Myc amplification.

Keywords: 4′,6-diamidino-2-phenylindole; APG; Apigenin; Apoptosis; BSA; CTL; DAPI; Differentiation; FITC; GAPDH; HRP; ICAD; ID2; MMP-2; N-Myc shRNA; NF-κB; NFP; NSE; Neuroblastoma; PBS; PCNA; PI; PVDF; RT-PCR; SBDP; SDS-PAGE; VEGF; apigenin; b-FGF; basic fibroblast growth factor; bovine serum albumin; control; fluorescein isothocyanate; glyceraldehyde 3-phosphate dehydrogenase; hTERT; horseradish peroxidase; human telomerase reverse transcriptase; inhibitor of caspase-activated DNase; inhibitor of differentiation 2; matrix metalloproteinase-2; neurofilament protein; neuron specific enolase; nuclear factor-kappa B; phosphate-buffered saline; polyvinylidene fluoride; proliferating cell nuclear antigen; propidium iodide; reverse transcription-polymerase chain reaction; shRNA; short hairpin RNA; sodium dodecyl sulfate-polyacrylamide gel electrophoresis; spectrin breakdown product; vascular endothelial growth factor.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Apigenin / pharmacology*
  • Apoptosis / drug effects
  • Caspase 3 / genetics
  • Caspase 3 / metabolism
  • Caspase 8 / genetics
  • Caspase 8 / metabolism
  • Cell Differentiation / drug effects
  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Gene Knockdown Techniques
  • Humans
  • Neuroblastoma / genetics*
  • Neuroblastoma / pathology
  • Proto-Oncogene Proteins c-myc / genetics*
  • Proto-Oncogene Proteins c-myc / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Transfection

Substances

  • Antineoplastic Agents
  • Proto-Oncogene Proteins c-myc
  • RNA, Messenger
  • RNA, Small Interfering
  • Apigenin
  • CASP3 protein, human
  • CASP8 protein, human
  • Caspase 3
  • Caspase 8