ORIGINAL ARTICLE

Comparison of gene-expression profiles in parallel bone marrow and peripheral blood samples in acute myeloid leukaemia by real-time polymerase chain reaction

E Sakhinia¹, M Farahangpour¹, E Tholouli², J A Liu Yin², J A Hoyland¹ and R J Byers¹

¹ Faculty of Medical and Human Sciences, Division of Laboratory and Regenerative Medicine, School of Medicine, The University of Manchester, Manchester, UK
² Department of Haematology, Manchester Royal Infirmary, Manchester

Correspondence to:
Correspondence to:
R J Byers
Division of Laboratory and Regenerative Medicine, The University of Manchester, Stopford Building, Oxford Road, Manchester M13 9PT, UK; r.byers{at}manchester.ac.uk

Background: Gene signatures (Indicator genes) in bone marrow that provide more precise prognostication in haematological malignancy have been identified by microarray expression studies. It would be beneficial to measure these diagnostic signatures in peripheral blood.

Aims: To determine the degree of correspondence of gene expression for a set of Indicator genes between bone marrow and peripheral blood in acute myeloid leukaemia (AML).

Methods: Parallel bone marrow aspirate and peripheral blood samples were obtained from 19 patients diagnosed with AML and mononuclear cells isolated from both sample types. mRNA was globally amplified by polyadenylated real-time polymerase chain reaction (polyA RT-PCR); the expression of 15 AML Indicator genes, identified from previous microarray studies, was measured by RT-PCR. All values were normalised to the mean expression of three housekeeping genes (IF2-ß, GAP and RbS9) and were statistically compared using SPSS software.

Results: No significant difference in expression between bone marrow and peripheral blood was observed for 10 of the genes (leptin receptor, CD33, adipsin, proteoglycan 1, MB-1, cyclin D3, hSNF2b, proteasome iota, HkrT-1 and E2A), indicating its possible use in monitoring disease activity in peripheral blood samples, whereas c-myb, HOXA9, LYN, cystatin c and LTC4s showed significantly different expression between bone marrow and peripheral blood samples.

Conclusion: These results indicate a possible use for the method in monitoring AML in peripheral blood by RT-PCR measurement of Indicator genes. In addition, the initial use of polyA PCR facilitates translation to very small clinical samples, including fractionated cell populations, of particular importance for monitoring haematological malignancy.

Abbreviations: AML, acute myeloid leukaemia; ALL, acute lymphoblastic leukaemia; Mhouse, mean of the three housekeeping genes; polyA PCR, polyadenylated polymerase chain reaction; RbS9, ribosomal protein S9; RT-PCR, real-time polymerase chain reaction

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