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.
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.
Niwa
K
K
Department of Obstetrics and Gynecology, Gifu University School of Medicine.
Misao
R
R
Hanabayashi
T
T
Morishita
S
S
Murase
T
T
Itoh
M
M
Itoh
N
N
Mori
H
H
Tamaya
T
T
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
D017931:DNA Primers; D004273:DNA, Neoplasm; D012333:RNA, Messenger; D012194:RNA-Directed DNA Polymerase; D004264:DNA Topoisomerases, Type I; D002166:Camptothecin