Vinogradov et al. (Abstract)
Cellular Therapy and Transplantation (CTT), Vol. 3, No. 9
doi: 10.3205/ctt-2010-No9-abstract05
© The Authors. This abstract is provided under the following license: Creative Commons Attribution 3.0 Unported
Abstract accepted for "4th Raisa Gorbacheva Memorial Meeting on Hematopoietic Stem Cell Transplantation",
Saint Petersburg, Russia, September 18–20, 2010
TP53 gene point mutation detection using sequencing techniques in acute and chronic leukemia patients
Alexander V. Vinogradov, Alexey V. Rezaikin, Alexander G. Sergeev
Ural State Medical Academy, Ekaterinburg, Russia
Correspondence: Alexander V. Vinogradov, Ural State Medical Academy, 3, Repina str., 620028, Ekaterinburg, Russia, E-mail: vinogradov-av@ya.ru
Abstract
Aim: TP53 gene point mutation detection in acute and chronic leukemia patients by using sequencing methods.
Patients and methods: 24 patients were included in the study, among which were 6 pts with AML, 9 with ALL, 6 with CML, 1 with idiopathic myelofibrosis, and 1 with aggressive lymphoma. TP53 gene point mutation detection was realized via the sequencing method. The extraction of mRNA was made from bone marrow material and peripheral blood neoplastic cells. Sequences of primers for TP53 gene amplification (exons 4–11) using a polymerase chain reaction technique are represented in table 1.
Table 1.
Primer | Sequence | Position |
|---|---|---|
P53-F | 5' AGT CAG ATC CTA GCG TCG AG 3' | 211–230 |
P53-R | 5' TGG CGG GAG GTA GAC TG 3' | 1320–1336 |
Primers for the termination reaction for amplified TP53 gene loci were chosen using highly conservative and specific mRNA nucleotide sequences represented in the GenBank (table 2).
Table 2.
Primer | Sequence | Position |
|---|---|---|
P53-S1 | 5' ACA TTT TCA GAC CTA TGG AA 3' | 249–268 |
P53-S2 | 5' GCG CCA TGG CCA TCT ACA AG 3' | 670–689 |
P53-S3 | 5' ACC CTT TTT GGA CTT CAG GT 3' | 1300–13 |
Sequencing was realized via an automatic genetic analyzer ABI Prism 310 using reactive components and an ABI Prism BigDye Terminator Cycle Sequencing Kit v. 3.0 (Applied Biosystem, USA).
Results: All patients from the research group except one with CLL showed the nucleotide change C215G (cytosine for guanine in position 215), which leads to a change of amino acid proline for arginine in position 72 (P72R) in the determining protein. In one case the variant of gene TP53 according to wild type (NM_000546, GenBank NCBI) was detected.
Keywords: TP53 gene, sequencing, leukemia, C215G nucleotide change
