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Blood, Vol. 94 No. 3 (August 1), 1999:
pp. 1113-1120
Loss of p73 Gene Expression in Leukemias/Lymphomas Due to
Hypermethylation
Seiji Kawano,
Carl W. Miller,
Adrian F. Gombart,
Claus R. Bartram,
Yoshinobu Matsuo,
Hiroya Asou,
Akiko Sakashita,
Jonathan Said,
Eiji Tatsumi, and
H. Phillip Koeffler
From Hematology/Oncology, Cedars-Sinai Medical Center, UCLA School of
Medicine, Los Angeles, CA; Institut fur Humangenetik,
Ruprecht-Karls-Universitat Heidelberg, Heidelberg, Germany; Fujisaki
Cell Center, Hayashibara Biological Laboratories Inc, Okayama, Japan;
the Department of Cancer Genetics, Research Institute for Radiation
Biology and Medicine, Hiroshima University, Hiroshima, Japan; the
Department of Hematology, Saitama Cancer Center, Saitama, Japan; the
Department of Pathology and Laboratory Medicine, UCLA School of
Medicine, Los Angeles, CA; and the International Center for Medical
Research, Kobe University School of Medicine, Kobe, Japan.
The p73 gene, a member of the p53 family, is a new
candidate tumor suppressor gene. To investigate the possibility of
genetic alteration of p73 in leukemia and lymphoma, we examined
55 cell lines and 39 patient samples together with 17 nonhematopoietic cancer cell lines. Gene expression of p73 was detected by
reverse transcriptase-polymerase chain reaction (RT-PCR) in cell lines (5 of 7 pre B/B-acute lymphoblastic leukemia [ALL], 13 of 21 T-ALL/lymphoblastic lymphomas [LBL], 9 of 10 B-non-Hodgkin's
lymphomas [B-NHL], 8 of 9 acute myelogenous leukemias [AML], 2 of 2 T-NHL, 3 of 3 multiple myeloma), and in patient samples (16 of 23 pre
B-ALL, 5 of 8 T-ALL/LBL, 5 of 8 B-NHL). PCR-single-strand conformation
polymorphism (SSCP) of cDNAs showed no mutation in 43 p73-expressing cell lines within the regions that corresponded
to the 5 mutational hotspots of the p53 gene. Neither
homologous deletion nor rearrangement of the p73 gene were
found by Southern blot analysis in any of the cell lines that lack
expression of p73. In contrast to prior published data,
analysis of a polymorphic site showed that the p73 gene was
expressed biallelically in cell lines and normal peripheral blood.
Notably, the p73-negative cell lines were hypermethylated at a
CpG island in the 5' untranslated region of the p73 mRNA, and treatment of these cell lines with 5-azacytidine (5-AC), a demethylation reagent, induced p73 expression. Taken together, we found that a sizable proportion (32%) of ALL/B-NHL cell lines and
primary tumors had negligible or limited expression of the p73
gene associated with hypermethylation of the gene. These findings suggest that silencing of the p73 gene by hypermethylation may contribute to development and/or progression of lymphoid neoplasms.
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