Open AccessSemi‐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
Author(s) -
Niwa Kenji,
Misao Ryou,
Hanabayashi Takabiro,
Morishita Shigeo,
Murase Toshiko,
Itoh Miho,
Itoh Naoki,
Mori Hidehiro,
Tamaya Teruhiko
Publication year - 1994
Publication title -
japanese journal of cancer research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.035
H-Index - 141
eISSN - 1349-7006
pISSN - 0910-5050
DOI - 10.1111/j.1349-7006.1994.tb02960.x
Subject(s) - microbiology and biotechnology , topoisomerase , camptothecin , biology , dna , transcription (linguistics) , reverse transcriptase , polymerase chain reaction , cytotoxicity , messenger rna , cell culture , polymerase , reverse transcription polymerase chain reaction , gene , biochemistry , genetics , in vitro , linguistics , philosophy
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.