title: Real-time RT-PCR for the determination of topoisomerase II mRNA level in leukaemic cells.
abstract: We developed a real-time RT-PCR assay for the quantification of topoisomerase II (topo II) mRNA level. It was applied on peripheral leukaemic cells from 23 patients with acute myelogenous leukaemia (AML) and 23 with chronic lymphocytic leukaemia (CLL). RNA template dilutions from 0.25 to 25ng per reaction were used as standard curves for topo IIalpha, beta and the internal control 18S rRNA. About 57% (26/46) and 26% (12/46) of the specimens had detectable topo IIbeta and alpha mRNA, respectively. The correlation between these two factors was rho=0.7 and P=0.0001. No relationship between topo IIalpha or beta mRNA level and response to chemotherapy was found in AML patients (n=19 assessable for response). Our method is rapid and convenient for quantification of topo IIalpha and beta mRNA levels, and could be suitable for investigation in a larger population.
authors.affiliation: Department of Oncology-Pathology, Cancer Center Karolinska at CCK, Karolinska Institute and Hospital, R8:00, SE-171 76 Stockholm, Sweden. rong.zhou@cck.ki.se
authors.forename: Rong
authors.initials: R
authors.lastname: Zhou
authors.forename: Marianne
authors.initials: M
authors.lastname: Frostvik Stolt
authors.forename: Ulrike
authors.initials: U
authors.lastname: Kronenwett
authors.forename: Astrid
authors.initials: A
authors.lastname: Gruber
authors.forename: Jan
authors.initials: J
authors.lastname: Liliemark
authors.forename: Eva
authors.initials: E
authors.lastname: Liliemark
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D004250:DNA Topoisomerases, Type II / Q000235:genetics*; D019008:Drug Resistance, Neoplasm; D005260:Female; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000188:drug therapy / Q000201:enzymology*; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy / Q000201:enzymology*; D008297:Male; D008875:Middle Aged; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
chemical_list: D012333:RNA, Messenger; D004250:DNA Topoisomerases, Type II