Quantitation of somatostatin receptor type 2 gene expression in neuroblastoma cell lines and primary tumors using competitive reverse transcription-polymerase chain reaction

Clin Cancer Res. 1996 Oct;2(10):1757-65.

Abstract

We previously reported the presence of somatostatin (SS-14)-binding sites in a wide panel of human neuroblastoma (NB) tumor cell lines. Given that the adrenal gland and its relative embryonal and adult tumors express an abundance of mRNA for somatostatin receptor type 2 (sst2) mRNA, we studied the quantitative expression of sst2 in 6 NB cell lines and 15 primary tumors using competitive reverse transcription (RT)-PCR. This method uses an insertion mutant of the target gene as a competitor for the RT-PCR reaction, thus allowing exact quantitation of sst2 mRNA abundance. We found expression of specific transcripts for sst2 in all of the NB cell lines and tumors investigated (range, 9 x 10(5)-4 x 10(9) molecules/microg RNA). In NB cells, the expression of sst2 was highly correlated with SS-14-binding sites (R = 0.93). In primary tumors, sst2 was positively related to the expression of the neuroendocrine marker secretogranin II (P < 0.05) and negatively related to N-myc amplification (a poor prognostic factor, P < 0.005) and metastatic dissemination (P < 0.05). In addition, Kaplan-Meier curves indicate that sst2 expression is positively related to survival (P = 0.01). In a patient with stage IVs disease (a spontaneously regressing form), we found the highest sst2 expression (4 x 10(9) molecules/microgram RNA), a value relatively similar to that of normal adrenal. In conclusion, these data indicate that quantitation of sst2, as assessed with competitive RT-PCR, could represent a new prognostic tool in the neuroendocrine tumor NB. Since sst2 recognizes octreotide with high affinity, these findings could also have both diagnostic and therapeutic value.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Binding Sites
  • Blotting, Northern
  • Child, Preschool
  • Female
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Infant
  • Infant, Newborn
  • Male
  • Neuroblastoma / genetics*
  • Neuroblastoma / pathology
  • Proto-Oncogene Proteins c-myc / genetics
  • RNA, Neoplasm / analysis
  • RNA, Neoplasm / genetics
  • Radioligand Assay
  • Receptors, Somatostatin / genetics*
  • Receptors, Somatostatin / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Somatostatin / metabolism
  • Survival Analysis
  • Transcription, Genetic
  • Tumor Cells, Cultured

Substances

  • Proto-Oncogene Proteins c-myc
  • RNA, Neoplasm
  • Receptors, Somatostatin
  • Somatostatin
  • somatostatin receptor 2