Reverse transcription polymerase chain reaction for the rearranged retinoic acid receptor alpha clarifies diagnosis and detects minimal residual disease in acute promyelocytic leukemia
W H Miller Jr, A Kakizuka, S R Frankel, R P Warrell Jr, A DeBlasio, K Levine, R M Evans, E Dmitrovsky, W H Miller Jr, A Kakizuka, S R Frankel, R P Warrell Jr, A DeBlasio, K Levine, R M Evans, E Dmitrovsky
Abstract
The characteristic t(15;17) of acute promyelocytic leukemia (APL) fuses the retinoic acid receptor alpha (RAR-alpha) gene on chromosome 17 to a gene on chromosome 15 called PML, a putative transcription factor. This distinct translocation results in a fusion mRNA detected by Northern analysis. Two cDNAs have been isolated that differ in the extent of 3' PML nucleic acid sequence contained. This study describes a reverse transcription polymerase chain reaction (RT-PCR) assay for the PML/RAR-alpha fusion transcript, which amplifies PML/RAR-alpha mRNA from APL cells with either reported breakpoint. DNA sequencing of the predominant RT-PCR products from 6 patients showed identical RAR-alpha exonic breakpoints and two PML breakpoints. This RT-PCR assay was positive in leukemic cells from 30/30 APL patients with the molecular rearrangement confirmed by cytogenetics or Northern analysis. In leukemic cells of patients with a morphologic diagnosis of APL lacking the t(15;17) by routine cytogenetics, a positive RT-PCR assay predicted clinical response to all-trans-retinoic acid (RA) therapy. Dilutional studies with leukemic cells that express (NB4) or do not express (HL-60) a PML/RAR-alpha fusion mRNA reveal that this RT-PCR assay detects the transcript from as little as 50 pg of total RNA. In APL cells from 5/6 patients treated with RA alone, a complete response by clinical and cytogenetic criteria accompanied a persistently positive RT-PCR assay. This preceded relapse by 1-6 months. RT-PCR for PML/RAR-alpha mRNA provides a more-sensitive test for the t(15;17) than routine cytogenetics or Northern analysis. This molecular rearrangement detected by RT-PCR best defines this RA-responsive malignancy. The RT-PCR assay for the PML/RAR-alpha transcript yields important diagnostic and prognostic information in the management of APL patients.
References
- Nouv Rev Fr Hematol. 1990;32(1):34-6
- Blood. 1990 Nov 1;76(9):1704-9
- Proc Natl Acad Sci U S A. 1988 Aug;85(15):5698-702
- Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7
- Oncogene. 1991 Jul;6(7):1285-92
- Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1977-81
- N Engl J Med. 1991 May 16;324(20):1385-93
- Blood. 1991 Mar 1;77(5):1080-6
- Leukemia. 1990 Dec;4(12):802-7
- J Exp Med. 1990 Dec 1;172(6):1571-5
- Science. 1990 Oct 26;250(4980):559-62
- Nature. 1987 Dec 3-9;330(6147):444-50
- Blood. 1988 Aug;72(2):567-72
- Blood. 1988 Jan;71(1):242-7
- Ann Intern Med. 1985 Oct;103(4):620-5
- Biochemistry. 1979 Nov 27;18(24):5294-9
- J Mol Biol. 1975 Nov 5;98(3):503-17
- Cell. 1991 Aug 23;66(4):663-74
- Cell. 1991 Aug 23;66(4):675-84
- Blood. 1991 Apr 1;77(7):1418-22
- Blood. 1991 Sep 15;78(6):1413-9
- Nucleic Acids Res. 1988 Nov 11;16(21):10366
- Cancer Genet Cytogenet. 1986 Feb 15;20(3-4):209-22
- Am J Med. 1984 May;76(5):827-41
- Blood. 1991 Feb 15;77(4):874-8
- Nature. 1990 Oct 11;347(6293):558-61
- J Natl Cancer Inst. 1990 Dec 19;82(24):1932-3
- Blood. 1990 Jun 1;75(11):2112-7
- Nature. 1987 Dec 17-23;330(6149):624-9
Source: PubMed