Real-time RT-PCR for the determination of topoisomerase II mRNA level in leukaemic cells.
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.
Zhou
Rong
R
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
Frostvik Stolt
Marianne
M
Kronenwett
Ulrike
U
Gruber
Astrid
A
Liliemark
Jan
J
Liliemark
Eva
E
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
D012333:RNA, Messenger; D004250:DNA Topoisomerases, Type II