Analysis of Acute Myelogenous Leukemia: Preparation of Samples for Genomic and Proteomic Analyses

Abstract

During the last decade, several large clinical studies have demonstrated that analysis of chromosomal abnormalities is an essential basis for therapeutic decisions in patients with acute myelogenous leukemia (AML), and cytogenetic studies should now be regarded as mandatory both for routine treatment and as a part of clinical investigations in AML. However, new techniques for detailed genetic characterization and analysis of gene expression as well as protein modulation will become important in the further classification of AML subsets and the development of risk-adapted therapeutic strategies. In this context, we emphasize the importance of population-based clinical studies as a basis for future therapeutic guidelines. Such studies will then require the inclusion of patients at small clinical centers without specialized hematological research laboratories. To document a high and uniform quality of the laboratory investigations, it will be necessary to collect material for later analysis in selected laboratories. In this article, we describe current methods for collection of biological samples that can be used for later preparation of DNA, RNA, and proteins. With the use of gradient-separated AML cells, it should be possible to establish the necessary techniques for collection and handling of biological samples even at smaller centers, and complete collections from all included patients should then be possible even in population-based clinical studies.

Correlation analysis of p53 protein isoforms with NPM1/FLT3 mutations and therapy response in acute myeloid leukemia

“…During Table 1. Cell separation, cryopreservation, culture and protein sample preparation of patient AML blasts and peripheral blood mononuclear cells from healthy donors were performed as previously described (Gjertsen et al, 2002;Bruserud et al, 2003;Irish et al, 2004). Presence of mRNA for p53 isoforms was examined by reverse transcriptase-PCR in approximately 50% of the patients according to previously described procedure (Bourdon et al, 2005).…”

Angiogenesis-independent tumor growth mediated by stem-like cancer cells

“…The Agilent 16,000-oligonucleotide cDNA microarrays were processed as described (53). ABI1700 DNA Oligonucleotide Microarrays.…”

A Distinct p53 Protein Isoform Signature Reflects the Onset of Induction Chemotherapy for Acute Myeloid Leukemia

“…Material for protein analysis was collected as described previously in detail (33,34). Briefly, cells were washed in NaCl (9 mg/mL) and then lysed in 7% TCA.…”