title: Isolation and characterization of novel RECK tumor suppressor gene splice variants.
abstract: Glioblastoma multiforme is the most common and lethal of the central nervous system glial-derived tumors. RECK suppresses tumor invasion by negatively regulating at least three members of the matrix metalloproteinase family: MMP-9, MMP-2, and MT1-MMP. A positive correlation has been observed between the abundance of RECK expression in tumor samples and a more favorable prognosis for patients with several types of tumors. In the present study, novel alternatively spliced variants of the RECK gene: RECK-B and RECK-I were isolated by RT-PCR and sequenced. The expression levels and profiles of these alternative RECK transcripts, as well as canonical RECK were determined in tissue samples of malignant astrocytomas of different grades and in a normal tissue RNA panel by qRT-PCR. Our results show that higher canonical RECK expression, accompanied by a higher canonical to alternative transcript expression ratio, positively correlates with higher overall survival rate after chemotherapeutic treatment of GBM patients. U87MG and T98G cells over-expressing the RECK-B alternative variant display higher anchorage-independent clonal growth and do not display modulation of, respectively, MMP-2 and MMP-9 expression. Our findings suggest that RECK transcript variants might have opposite roles in GBM biology and the ratio of their expression levels may be informative for the prognostic outcome of GBM patients.
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, 05508-000, Brazil.|NUCEL-NETCEM-Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, 05360-120, Brazil.
authors.forename: Marina
authors.initials: M
authors.lastname: Trombetta-Lima
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Curitiba, PR, 81531-990, Brazil.
authors.forename: Sheila Maria Brochado
authors.initials: SM
authors.lastname: Winnischofer
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, 05508-000, Brazil.|NUCEL-NETCEM-Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, 05360-120, Brazil.
authors.forename: Marcos Angelo Almeida
authors.initials: MA
authors.lastname: Demasi
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, 05508-000, Brazil.|NUCEL-NETCEM-Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, 05360-120, Brazil.
authors.forename: Renato
authors.initials: R
authors.lastname: Astorino Filho
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, SP, 05508-000, Brazil.|NUCEL-NETCEM-Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, 05360-120, Brazil.
authors.forename: Ana Claudia Oliveira
authors.initials: AC
authors.lastname: Carreira
authors.affiliation: Radiation Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892-4605, USA.
authors.forename: Beiyang
authors.initials: B
authors.lastname: Wei
authors.affiliation: NUCEL-NETCEM-Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, 05360-120, Brazil.
authors.forename: Thais
authors.initials: T
authors.lastname: de Assis-Ribas
authors.affiliation: NUCEL-NETCEM-Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, 05360-120, Brazil.
authors.forename: Michelle Silberspitz
authors.initials: MS
authors.lastname: Konig
authors.affiliation: NUCEL-NETCEM-Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, 05360-120, Brazil.|Dana Farber Cancer Institute, Harvard Medical School, Cambridge, MA, 02138, USA.
authors.forename: Christian
authors.initials: C
authors.lastname: Bowman-Colin
authors.affiliation: NUCEL-NETCEM-Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, 05360-120, Brazil.|Departmento de Neurologia, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, 01246-000, Brazil.
authors.forename: Sueli Mieko
authors.initials: SM
authors.lastname: Oba-Shinjo
authors.affiliation: NUCEL-NETCEM-Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, 05360-120, Brazil.|Departmento de Neurologia, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, 01246-000, Brazil.
authors.forename: Suely Kazue Nagahashi
authors.initials: SK
authors.lastname: Marie
authors.affiliation: Radiation Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892-4605, USA.
authors.forename: William
authors.initials: W
authors.lastname: Stetler-Stevenson
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo 05508-000 SP, Brazil.|NUCEL-NETCEM-Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, 05360-120, Brazil.
authors.forename: Mari Cleide
authors.initials: MC
authors.lastname: Sogayar
mesh_terms: D000328:Adult; D001932:Brain Neoplasms / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D005260:Female; D058851:GPI-Linked Proteins / Q000235:genetics* / Q000378:metabolism; D016147:Genes, Tumor Suppressor; D005909:Glioblastoma / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008297:Male; D020782:Matrix Metalloproteinases / Q000235:genetics / Q000378:metabolism; D008875:Middle Aged; D020033:Protein Isoforms; D012326:RNA Splicing
chemical_list: D058851:GPI-Linked Proteins; D020033:Protein Isoforms; C115460:RECK protein, human; D020782:Matrix Metalloproteinases
keywords: GBM; MMP; RECK; isoforms; splicing