Spermidine/spermine N1-acetyltransferase (SSAT) activity in human small-cell lung carcinoma cells following transfection with a genomic SSAT construct

Biochem J. 2003 Jul 15;373(Pt 2):629-34. doi: 10.1042/BJ20021895.

Abstract

Spermidine/spermine N (1)-acetyltransferase (SSAT) activity is typically highly inducible in non-small-cell lung carcinomas in response to treatment with anti-tumour polyamine analogues, and this induction is associated with subsequent cell death. In contrast, cells of the small-cell lung carcinoma (SCLC) phenotype generally do not respond to these compounds with an increase in SSAT activity, and usually are only moderately affected with respect to growth. The goal of the present study was to produce an SSAT-overexpressing SCLC cell line to further investigate the role of SSAT in response to these anti-tumour analogues. To accomplish this, NCI-H82 SCLC cells were stably transfected with plasmids containing either the SSAT genomic sequence or the corresponding cDNA sequence. Individual clones were selected based on their ability to show induced SSAT activity in response to exposure to a polyamine analogue, and an increase in the steady-state SSAT mRNA level. Cells transfected with the genomic sequence exhibited a significant increase in basal SSAT mRNA expression, as well as enhanced SSAT activity, intracellular polyamine pool depletion and growth inhibition following treatment with the analogue N (1), N (11)-bis(ethyl)norspermine. Cells containing the transfected cDNA also exhibited an increase in the basal SSAT mRNA level, but remained phenotypically similar to vector control cells with respect to their response to analogue exposure. These studies indicate that both the genomic SSAT sequence and polyamine analogue exposure play a role in the transcriptional and post-transcriptional regulation and subsequent induction of SSAT activity in these cells. Furthermore, this is the first production of a cell line capable of SSAT protein induction from a generally unresponsive parent line.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acetyltransferases / genetics*
  • Acetyltransferases / metabolism*
  • Blotting, Northern
  • Blotting, Southern
  • Blotting, Western
  • Carcinoma, Small Cell / enzymology*
  • Carcinoma, Small Cell / genetics
  • Carcinoma, Small Cell / pathology
  • Cell Division / drug effects
  • DNA, Complementary
  • Dose-Response Relationship, Drug
  • Enzyme Induction
  • Gene Expression Regulation, Enzymologic*
  • Humans
  • Lung Neoplasms / enzymology*
  • Lung Neoplasms / genetics
  • Lung Neoplasms / pathology
  • Protein Biosynthesis
  • RNA Processing, Post-Transcriptional
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Neoplasm
  • Transcription, Genetic
  • Transfection
  • Tumor Cells, Cultured

Substances

  • DNA, Complementary
  • RNA, Messenger
  • RNA, Neoplasm
  • Acetyltransferases
  • diamine N-acetyltransferase