title: Novel CT domain-encoding splice forms of CTGF/CCN2 are expressed in B-lineage acute lymphoblastic leukaemia.
abstract: BACKGROUND
  Connective tissue growth factor (CTGF/CCN2) has been shown previously to be aberrantly expressed in a high proportion of paediatric precursor B cell acute lymphoblastic leukaemia (pre-B ALL), suggesting a potential oncogenic role in this tumour type. We therefore assessed CTGF mRNA transcript diversity in B-lineage ALL using primary patient specimens and cell lines.
  
  
  METHODS
  CTGF mRNA expression was evaluated by quantitative real-time PCR and Northern blotting. We performed a structural analysis of CTGF mRNA by nested reverse-transcriptase PCR and examined CTGF protein diversity by immunoblotting.
  
  
  RESULTS
  Northern blot analysis of pre-B ALL cell lines revealed short CTGF transcripts that were expressed in association with the active phase of cellular growth. Structural analysis confirmed the synthesis of several novel CTGF mRNA isoforms in B-lineage ALL cell lines that were uniformly characterised by the retention of the coding sequence for the C-terminal (CT) domain. One of these novel spliceforms was expressed in a majority (70%) of primary pre-B ALL patient specimens positive for canonical CTGF mRNA. Evidence that these alternative transcripts have coding potential was provided by cryptic CTGF proteins of predicted size detected by immunoblotting.
  
  
  CONCLUSIONS
  This study identifies for the first time alternative splicing of the CTGF gene and shows that a short CTGF splice variant associated with cell proliferation is expressed in most cases of primary CTGF-positive pre-B ALL. This novel variant encoding only the CT domain may play a role in pre-B ALL tumorigenesis and/or progression.
authors.affiliation: Division of Children's Leukemia and Cancer Research, Telethon Kids Institute, The University of Western Australia, Perth, WA, Australia; Faculty of Health Sciences, School of Pharmacy, Curtin University, Perth, WA, Australia. Electronic address: mat.welch@uwa.edu.au.
authors.forename: M D
authors.initials: MD
authors.lastname: Welch
authors.affiliation: Division of Children's Leukemia and Cancer Research, Telethon Kids Institute, The University of Western Australia, Perth, WA, Australia. Electronic address: meegan.howlett@telethonkids.org.au.
authors.forename: M
authors.initials: M
authors.lastname: Howlett
authors.affiliation: Division of Children's Leukemia and Cancer Research, Telethon Kids Institute, The University of Western Australia, Perth, WA, Australia. Electronic address: heloise.halse@telethonkids.org.au.
authors.forename: H M
authors.initials: HM
authors.lastname: Halse
authors.affiliation: School of Veterinary and Life Sciences, Murdoch University, Perth, WA, Australia. Electronic address: w.greene@murdoch.edu.au.
authors.forename: W K
authors.initials: WK
authors.lastname: Greene
authors.affiliation: Division of Children's Leukemia and Cancer Research, Telethon Kids Institute, The University of Western Australia, Perth, WA, Australia. Electronic address: ursula.kees@telethonkids.org.au.
authors.forename: U R
authors.initials: UR
authors.lastname: Kees
mesh_terms: D002471:Cell Transformation, Neoplastic / Q000235:genetics; D002648:Child; D055513:Connective Tissue Growth Factor / Q000737:chemistry / Q000235:genetics*; D018450:Disease Progression; D020869:Gene Expression Profiling; D015973:Gene Expression Regulation, Leukemic; D006801:Humans; D015452:Precursor B-Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics*; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D014407:Tumor Cells, Cultured
chemical_list: C526317:CCN2 protein, human; D020033:Protein Isoforms; D055513:Connective Tissue Growth Factor
keywords: Acute lymphoblastic leukaemia; Alternative splicing; CCN2; CTGF; Pre-B ALL