title: Molecular heterogeneity in the novel fusion geneAPIP-FGFR2: Diversity of genomic breakpoints in gastric cancer with high-level amplifications at 11p13 and 10q26.
abstract: Several novel fusion transcripts were identified by next-generation sequencing in gastric cancer; however, the breakpoint junctions have yet to be characterized. The present study characterized a plethora ofAPIP-FGFR2genomic breakpoints in the SNU-16 gastric cancer cell line, which harbored homogeneously staining regions (hsrs) and double minute chromosomes. Oligonucleotide microarrays revealed high-level amplifications at chromosomes 8q24.1 (0.8 Mb region), 10q26 (1.1 Mb) and 11p13 (1.1 Mb). These amplicons containedMYCandPVT1at chromosome 8q24.1,BRWD2,FGFR2andATE1at chromosome 10q26, and 24 genes, includingAPIP,CD44,RAG1andRAG2, at chromosome 11p13. Based on these findings, reverse transcription-polymerase chain reaction (PCR) was performed using various candidate gene primers to detect possible fusion transcripts, and several products using primer sets for theAPIPandFGFR2genes were detected. Eventually, three in-frame and two out-of-frame fusion transcripts were detected. Notably, PCR analysis of the entire genomic DNA detected three distinct genomic junctions. The breakpoints were within intron 5 ofAPIP, which contained three distinct breakpoints, and introns 5, 7 and 9 ofFGFR2. Fluorescencein situhybridization showed several fusion signals within hsrs using two short probes (~10-kb segments of a bacterial artificial chromosome clone) containing exons 2-5 ofAPIPor exons 11-13 ofFGFR2. Although, for any given fusion, a multiplicity of transcripts is thought to be created by alternative splicing of one rearranged allele, the results of the present study suggested that genomic fusions ofAPIPandFGFR2are generated in hsrs with a diversity of breakpoints that are then faithfully transcribed.
authors.affiliation: Department of Gastroenterology and Hepatology, Kyoto Prefectural University of Medicine Graduate School of Medical Science, Kyoto 602-8566, Japan; Department of Hematology and Oncology, Kyoto Prefectural University of Medicine Graduate School of Medical Science, Kyoto 602-8566, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Okuda
authors.affiliation: Department of Molecular Diagnostics and Therapeutics, Kyoto Prefectural University of Medicine Graduate School of Medical Science, Kyoto 602-8566, Japan.
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Taki
authors.affiliation: Department of Hematology and Oncology, Kyoto Prefectural University of Medicine Graduate School of Medical Science, Kyoto 602-8566, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Nishida
authors.affiliation: Department of Hematology and Oncology, Kyoto Prefectural University of Medicine Graduate School of Medical Science, Kyoto 602-8566, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Chinen
authors.affiliation: Department of Hematology and Oncology, Kyoto Prefectural University of Medicine Graduate School of Medical Science, Kyoto 602-8566, Japan.
authors.forename: Hisao
authors.initials: H
authors.lastname: Nagoshi
authors.affiliation: Department of Digestive Surgery, Kyoto Prefectural University of Medicine Graduate School of Medical Science, Kyoto 602-8566, Japan.
authors.forename: Chouhei
authors.initials: C
authors.lastname: Sakakura
authors.affiliation: Department of Hematology and Oncology, Kyoto Prefectural University of Medicine Graduate School of Medical Science, Kyoto 602-8566, Japan.
authors.forename: Masafumi
authors.initials: M
authors.lastname: Taniwaki
keywords: APIP; FGFR2; gastric cancer; genomic amplification; homogenously staining region