|Budget Amount *help
¥2,300,000 (Direct Cost : ¥2,300,000)
Fiscal Year 1996 : ¥900,000 (Direct Cost : ¥900,000)
Fiscal Year 1995 : ¥1,400,000 (Direct Cost : ¥1,400,000)
1.Southern blot analysis using probes from the major translocation cluster region of the BCL6 revealed rearrangement in 21 of 197 patients (10.7%) with B-cell neoplasms who were studied at the time of presentation, and 11 of 25 patients (44%) first studied at relapse. BCL6 gene rearrangement was primarily involved in large cell lymphoma irrespective of growth pattern of neoplastic cells, and non-Hodgkin's lymphoma (NHL) with this lesion could be curable with modern intensive chemotherapy.
2.3q27 translocations affecting the BCL6 can involve not only immunoglobulin genes but also other as yet uncharacterized chromosomal loci as partners. We cloned junctional areas of a recurring translocation, t (3 ; 6) (q27 ; p21). Nucleotide sequence analysis of a fragment from the junctional area of 6p21.3 revealed the presence of a novel H4 histone gene. Breakpoints on 3q27 of two cases carrying the t (3 ; 6) were immediately 3' of the BCL6 exon 1, and the H4 histone gene was substituted for the 5' r
egulatory elements of the BCL6. Because H4 gene expression is tightly coupled to DNA replication, this study suggested an immediate mechanism for deregulated expression of the BCL6 leading to the development of NHL.
3.To establish a rapid and sensitive method to detect neoplastic cells carrying a specific chromosomal translocation, we developed a novel strategy based on long distance polymerase chain reaction (LD-PCR) amplification. LD-PCR with a newly available Taq polymerase for longer product synthesis successfully amplified fragments encompassing junctions of t (14 ; 18), t (8 ; 14), t (3 ; 14), t (3 ; 22), t (2 ; 3), t (14 ; 19) and t (9 ; 14). The sizes of the amplified fragments ranged from 2 kb to 30 kb, and the sensitivity was 10^<-4>. LD-PCR for t (14 ; 18) totally substituted for time-consuming Southern blot analysis. t (8 ; 14) was detectable in biopsies as well as postmorten materials. LD-PCR provided a useful tool for diagnosis and subsequent management of B-cell neoplasms characterized by specific chromosomal translocations.
3、染色体転座の転座接合部を効率よく検出する新しいPCR増幅法(Long distance PCR, LD-PCR)を開発した。B細胞性腫瘍に特異的な染色体転座であるt (14 ; 18), t (8 ; 14), t (3 ; 14), t (3 ; 22) , t (2 ; 3) , t (14 ; 19) , t (9 ; 14)の転座接合部を含む2-30kbのDNA断片の増幅が可能であり、感度は1/10^4である。t (14 ; 18) のLD-PCRによる検出は、従来のサザンブロットに完全にとってかわるものである。t (8 ; 14)のLD-PCRによる検出は、臨床検体や剖検検体に応用し、その有用性を明らかにした。 Less