Proteomic analysis of hematopoietic stem cell-like fractions in leukemic disorders.
DNA microarray analysis has been applied to identify molecular markers of human hematological malignancies. However, the relatively low correlation between the abundance of a given mRNA and that of the encoded protein makes it important to characterize the protein profile directly, or 'proteome,' of malignant cells in addition to the 'transcriptome.' To identify proteins specifically expressed in leukemias, here we isolated AC133(+) hematopoietic stem cell-like fractions from the bone marrow of 13 individuals with various leukemic disorders, and compared their protein profiles by two-dimensional electrophoresis. A total of 11 differentially expressed protein spots corresponding to 10 independent proteins were detected, and peptide fingerprinting combined with mass spectrometry of these proteins revealed them to include NuMA (nuclear protein that associates with the mitotic apparatus), heat shock proteins, and redox regulators. The abundance of NuMA in the leukemic blasts was significantly related to the presence of complex karyotype anomalies. Conditional expression of NuMA in a mouse myeloid cell line resulted in the induction of aneuploidy, cell cycle arrest in G(2)-M phases, and apoptosis. These results demonstrate the potential of proteome analysis with background-matched cell fractions obtained from fresh clinical specimens to provide insight into the mechanism of human leukemogenesis.
Ota
Jun
J
Division of Functional Genomics, Jichi Medical School, 3311-1 Yakushiji, Kawachi-gun, Tochigi 329-0498, Japan.
Yamashita
Yoshihiro
Y
Okawa
Katsuya
K
Kisanuki
Hiroyuki
H
Fujiwara
Shin-Ichiro
S
Ishikawa
Madoka
M
Lim Choi
Young
Y
Ueno
Shuichi
S
Ohki
Ruri
R
Koinuma
Koji
K
Wada
Tomoaki
T
Compton
Duane
D
Kadoya
Toshihiko
T
Mano
Hiroyuki
H
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000782:Aneuploidy; D000818:Animals; D034961:Antigens, Nuclear; D017209:Apoptosis; D018797:Cell Cycle Proteins; D005260:Female; D006412:Hematopoietic Stem Cells / Q000737:chemistry*; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000032:analysis*; D034521:Nuclear Matrix-Associated Proteins; D009687:Nuclear Proteins / Q000032:analysis; D040901:Proteomics; D015032:Zinc / Q000494:pharmacology
D034961:Antigens, Nuclear; D018797:Cell Cycle Proteins; C050045:NUMA1 protein, human; D009363:Neoplasm Proteins; D034521:Nuclear Matrix-Associated Proteins; D009687:Nuclear Proteins; C489080:Numa1 protein, mouse; D015032:Zinc