title: 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.
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.
authors.affiliation: Department of Obstetrics and Gynecology, Gifu University School of Medicine.
authors.forename: K
authors.initials: K
authors.lastname: Niwa
authors.forename: R
authors.initials: R
authors.lastname: Misao
authors.forename: T
authors.initials: T
authors.lastname: Hanabayashi
authors.forename: S
authors.initials: S
authors.lastname: Morishita
authors.forename: T
authors.initials: T
authors.lastname: Murase
authors.forename: M
authors.initials: M
authors.lastname: Itoh
authors.forename: N
authors.initials: N
authors.lastname: Itoh
authors.forename: H
authors.initials: H
authors.lastname: Mori
authors.forename: T
authors.initials: T
authors.lastname: Tamaya
mesh_terms: D000230:Adenocarcinoma; D001483:Base Sequence; D015139:Blotting, Southern; D002166:Camptothecin / Q000031:analogs & derivatives* / Q000633:toxicity*; D002460:Cell Line; D017931:DNA Primers; D004264:DNA Topoisomerases, Type I / Q000096:biosynthesis*; D004273:DNA, Neoplasm / Q000032:analysis; D016889:Endometrial Neoplasms; D005260:Female; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010051:Ovarian Neoplasms; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis*; D012194:RNA-Directed DNA Polymerase; D015183:Restriction Mapping; D014407:Tumor Cells, Cultured
chemical_list: D017931:DNA Primers; D004273:DNA, Neoplasm; D012333:RNA, Messenger; D012194:RNA-Directed DNA Polymerase; D004264:DNA Topoisomerases, Type I; D002166:Camptothecin