title: Rab15 alternative splicing is altered in spheres of neuroblastoma cells.
abstract: Neuroblastoma is an aggressive pediatric tumor that accounts for 15% of cancer-related deaths in children. More than half of high-risk neuroblastoma patients develop tumor relapse that is lethal in most cases. A small population of tumor-initiating cells (TICs), recently identified from high-risk neuroblastoma patients as spheres, is believed to be responsible for tumor relapse. Rab family small G proteins are essential in controlling membrane traffic and their misregulation results in several cancers. Rab15 was originally isolated as a brain-specific Rab protein regulating the endocytic recycling pathway and was recently identified as a downstream target of the neural transcription factor Atoh1. Previously, we identified two alternatively spliced Rab15 isoforms in neuroblastoma cells and showed a significant correlation between Rab15 expression and neuronal differentiation. As aberrant alternative splicing is intimately associated with an increasing number of cancers, its use as a new diagnostic and/or prognostic biomarker has attracted considerable attention. In the present study, we explored cancer-associated changes of Rab15 alternative splicing in neuroblastoma TICs. We found that Rab15 alternative splicing generated two novel isoforms designated as Rab15(AN2) and Rab15(AN3) in addition to two known isoforms designated as Rab15(CN) and Rab15(AN1). Although both Rab15(AN2) and Rab15(AN3) contained premature termination codons, they were detected in not only neuroblastoma cells but also in normal human tissues. One isoform was predominantly expressed in the brain and testis, while the other isoform was more specifically expressed in the brain. In neuroblastoma, Rab15 isoform balance measured by the Rab15(CN)/Rab15(AN1+AN2+AN3) ratio was significantly decreased in spheres compared to parental cells. These results suggest that Rab15 alternative splicing may serve as a biomarker to discriminate TICs from non-TICs in neuroblastoma.
authors.affiliation: Department of Pediatrics, Kobe University Graduate School of                     Medicine, Kobe, Japan.
authors.forename: Thi Van Huyen
authors.initials: TV
authors.lastname: Pham
authors.forename: Tri Budi
authors.initials: TB
authors.lastname: Hartomo
authors.forename: Myeong Jin
authors.initials: MJ
authors.lastname: Lee
authors.forename: Daiichiro
authors.initials: D
authors.lastname: Hasegawa
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Ishida
authors.forename: Keiichiro
authors.initials: K
authors.lastname: Kawasaki
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Kosaka
authors.forename: Tomoto
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Satoru
authors.initials: S
authors.lastname: Morikawa
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Yamamoto
authors.forename: Ikuko
authors.initials: I
authors.lastname: Kubokawa
authors.forename: Takeshi
authors.initials: T
authors.lastname: Mori
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yanai
authors.forename: Akira
authors.initials: A
authors.lastname: Hayakawa
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Takeshima
authors.forename: Kazumoto
authors.initials: K
authors.lastname: Iijima
authors.forename: Masafumi
authors.initials: M
authors.lastname: Matsuo
authors.forename: Hisahide
authors.initials: H
authors.lastname: Nishio
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Nishimura
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics*; D017398:Alternative Splicing / Q000235:genetics*; D014408:Biomarkers, Tumor / Q000235:genetics; D001921:Brain / Q000473:pathology; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D009447:Neuroblastoma / Q000235:genetics* / Q000473:pathology; D020033:Protein Isoforms / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D020691:rab GTP-Binding Proteins / Q000235:genetics
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D014408:Biomarkers, Tumor; D008565:Membrane Proteins; D020033:Protein Isoforms; D012333:RNA, Messenger; C509340:Rab15 effector protein, human; D020691:rab GTP-Binding Proteins