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[Cancer Research 60, 2335-2341, May 1, 2000]
© 2000 American Association for Cancer Research

Advances in Brief

Molecular Anatomy of BCL6 Translocations Revealed by Long-Distance Polymerase Chain Reaction-based Assays1

Hiroshi Akasaka2, Takashi Akasaka2, Masayuki Kurata, Chiyoko Ueda, Akira Shimizu, Takashi Uchiyama and Hitoshi Ohno3

First Division, Department of Internal Medicine, Faculty of Medicine [H. A., T. A., M. K., C. U., T. U., H. O.], and Center for Molecular Biology and Genetics [T. A., A. S.], Kyoto University, Kyoto 606-8507, Japan

BCL6 translocations involve not only immunoglobulin (IG) genes but also a number of non-IG loci as partners. Junctional sequences of three IG/BCL6 translocations were readily obtained by long-distance PCR. In cases where partner loci were not determined, we developed a long-distance inverse PCR method, which amplifies unknown fragments flanked by known BCL6 sequences. Using these two long-distance PCR-based approaches, we cloned junctional areas of BCL6 translocations from a total of 58 cases of B-cell tumors. Nucleotide sequencing and database searches revealed that 30 cases involved IGs as partners: IG heavy chain gene in 22, IG {kappa} light chain gene in 1, and IG {lambda} light chain gene in 7. In contrast, 23 cases affected non-IG loci, including the H4 histone gene, heat shock protein genes HSP89{alpha} and HSP90ß, and PIM-1 proto-oncogene. On der(3) chromosomes, complete sets of the promoters of these partner genes replaced that of BCL6 in the same transcriptional orientation. These results suggest that BCL6 gene affected by the translocation is transcriptionally activated by a variety of stimuli, including cell cycle control, changes in the physical environment, and response to cytokines. Break points on BCL6 occurred within the major translocation cluster, and we identified a 120-bp hyper-cluster region a short distance from the 3' end of exon 1. Gel mobility-shift assay suggested the presence of a protein(s) that bound to this particular region.




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Copyright © 2000 by the American Association for Cancer Research.