A PCR-aided transcript titration assay (PATTY) to measure topoisomerase I gene expression in human tumor specimens

GJ Meersma, M Bakker, HJM Groen, AGJ Van der Zee, PB Jensen, G Giaccone, EGE De Vries, EF Smit*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

6 Citations (Scopus)

Abstract

Topoisomerase I (topo I) inhibitors are promising anticancer agents with demonstrated activity against a wide range of solid tumors. Quantitative information on topol mRNA levels in tumor biopsies may predict response to topo I inhibitors. Materials and methods: A polymerase chain reaction aided transcript titration assay (PATTY) was developed to allow quantitation of topol mRNA in small samples. Concentrations of topol mRNA in total RNA samples were estimated by RT-PCR analysis in a human small cell lung cancer (SCLC) cell line (GLC,) and its topotecan (GL(2)C/SK and F) and camptothecin (GL(2)C/Campt) resistant sublines, human nonsmall cell lung cancer (NSCLC) and ovarian carcinoma samples. Results: Topol PATTY showed a decreased topo I mRNA level in GLC(2)/SK and F (4.5 pg/100 ng total RNA) and GLC,/Campt (2.2 pg/100 ng total RNA), respectively, compared to the parent cell line GLC(2) (5.4 pg/100 ng total RNA). Topol protein levels as measured by Western blotting were compatible with topol mRNA levels. Median (range) topol mRNA levels were 3.23 (2.33-5.10) pg/100 ng total RNA in resected NSCLC specimen (n=6), and 2.03 (0.54-0.95) pg/100 ng total RNA in resected ovarian cancer specimen (n=6). Conclusion: We conclude that topol PATTY is a new assay that quantitates topol mRNA levels in cell lines and small tumor samples.

Original languageEnglish
Pages (from-to)1309-1315
Number of pages7
JournalAnticancer Research
Volume19
Issue number2A
Publication statusPublished - 1999

Keywords

  • topoisomerase I gene expression
  • human tumor specimen
  • POLYMERASE CHAIN-REACTION
  • CANCER CELL-LINE
  • LUNG-CANCER
  • CATALYTIC ACTIVITY
  • RIBONUCLEIC-ACID
  • MESSENGER-RNA
  • COLON-CANCER
  • CAMPTOTHECIN
  • RESISTANT
  • CHEMOTHERAPY

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