Semi-quantitative analysis of DNA topoisomerase-I mRNA level using reverse transcription-polymerase chain reaction in cancer cell lines: its relation to cytotoxicity against camptothecin derivative

Jpn J Cancer Res. 1994 Aug;85(8):869-74. doi: 10.1111/j.1349-7006.1994.tb02960.x.

Abstract

Expression of DNA topoisomerase (Topo)-I-mRNA in various cancer cell lines was detected using the reverse transcription-polymerase chain reaction (RT-PCR) method. The cytoplasmic polyadenylated RNA isolated from cancer cell lines was reverse-transcribed and the complementary DNA was amplified by PCR primed with Topo-I specific primers. Fidelity of the amplified sequence was confirmed by restriction endonuclease digestion and Southern blot hybridization. The level of Topo-I mRNA was correlated positively with the cytotoxicity of a Topo-I inhibitor, a camptothecin derivative. This RT-PCR method may be applicable to the assessment of sensitivity of cells to Topo-I targeted drugs, especially when only small quantities of cell samples are available.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma
  • Base Sequence
  • Blotting, Southern
  • Camptothecin / analogs & derivatives*
  • Camptothecin / toxicity*
  • Cell Line
  • DNA Primers
  • DNA Topoisomerases, Type I / biosynthesis*
  • DNA, Neoplasm / analysis
  • Endometrial Neoplasms
  • Female
  • Gene Expression
  • HeLa Cells
  • Humans
  • Molecular Sequence Data
  • Ovarian Neoplasms
  • Polymerase Chain Reaction / methods*
  • RNA, Messenger / analysis*
  • RNA-Directed DNA Polymerase
  • Restriction Mapping
  • Tumor Cells, Cultured

Substances

  • DNA Primers
  • DNA, Neoplasm
  • RNA, Messenger
  • RNA-Directed DNA Polymerase
  • DNA Topoisomerases, Type I
  • Camptothecin