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1.
Cancer Res ; 61(21): 7978-84, 2001 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-11691822

RESUMO

A second adenomatous polyposis coli (APC)-like gene, APC2/APCL, was recently described and localized to chromosome 19. We have fine mapped APC2 to a small region of chromosome 19p13.3 containing markers D19S883 and WI-19632, a region commonly lost in a variety of cancers, particularly ovarian cancer. Interphase fluorescence in situ hybridization analysis revealed an APC2 allelic imbalance in 19 of 20 ovarian cancers screened and indicates that APC2 could be a potential tumor suppressor gene in ovarian cancer. When overexpressed in SKOV3 ovarian cancer cells, which express low levels of APC2, exogenous APC2 localized to the Golgi apparatus, actin-containing structures, and occasionally to microtubules. Antibodies against the NH2 terminus of human APC2 show that endogenous APC2 is diffusely distributed in the cytoplasm and colocalizes with both the Golgi apparatus and actin filaments. APC2 remained associated with actin filaments after treatment with the actin-disrupting agent, cytochalasin D. These results suggest that APC2 is involved in actin-associated events and could influence cell motility or adhesion through interaction with actin filaments, as well as functioning independently or in cooperation with APC to down-regulate beta-catenin signaling.


Assuntos
Desequilíbrio Alélico , Proteínas do Citoesqueleto/genética , Proteínas do Citoesqueleto/metabolismo , Citoesqueleto de Actina/metabolismo , Animais , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Cromossomos Humanos Par 19/genética , Proteínas do Citoesqueleto/biossíntese , Cães , Feminino , Expressão Gênica , Genes APC , Genes Supressores de Tumor , Complexo de Golgi/metabolismo , Humanos , Hibridização in Situ Fluorescente , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo , Mapeamento de Híbridos Radioativos , Transfecção , Células Tumorais Cultivadas
2.
J Biol Chem ; 275(45): 35478-85, 2000 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-10956646

RESUMO

An expression cloning approach was employed to identify the receptor for B-lymphocyte stimulator (BLyS) and identified the tumor necrosis factor receptor superfamily member TACI as a BLyS-binding protein. Expression of TACI in HEK293T cells confers on the cells the ability to bind BLyS with subnanomolar affinity. Furthermore, a TACI-Fc fusion protein recognizes both the cleaved, soluble form of BLyS as well as the membrane BLyS present on the cell surface of a recombinant cell line. TACI mRNA is found predominantly in B-cells and correlates with BLyS binding in a panel of B-cell lines. We also demonstrate that TACI interacts with nanomolar affinity with the BLyS-related tumor necrosis factor homologue APRIL for which no clear in vivo role has been described. BLyS and APRIL are capable of signaling through TACI to mediate NF-kappaB responses in HEK293 cells. We conclude that TACI is a receptor for BLyS and APRIL and discuss the implications for B-cell biology.


Assuntos
Linfócitos B/fisiologia , Proteínas de Membrana , Neuropeptídeos/fisiologia , Proteínas Nucleares/fisiologia , Receptores do Fator de Necrose Tumoral/fisiologia , Fator de Necrose Tumoral alfa/química , Receptor do Fator Ativador de Células B , Linfócitos B/metabolismo , Linhagem Celular , Membrana Celular/metabolismo , DNA Complementar/metabolismo , Relação Dose-Resposta a Droga , Citometria de Fluxo , Biblioteca Gênica , Humanos , Cinética , Ligantes , Reação em Cadeia da Polimerase , Ligação Proteica , RNA Mensageiro/metabolismo , Receptores do Fator de Necrose Tumoral/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Fatores de Tempo , Transfecção , Proteína Transmembrana Ativadora e Interagente do CAML
3.
Int J Oncol ; 13(5): 1061-7, 1998 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-9772300

RESUMO

Ligands of the EGF/Heregulin family control the growth of epithelial cells by binding to receptors of the erbB family. By searching a large database of cDNA sequences at Human Genome Sciences Inc. we have identified a new encoded protein sequence containing all the conserved elements of the EGF/Heregulin family. The same sequence has recently been independently identified as NRG-3. The EGF-like domain of NRG-3 was generated as a recombinant protein in E. coli and used to test the specificity of receptor binding. In human breast cancer cells and in 32D cells transfected by erbB family members, NRG-3 activated multiple erbB family members. These include EGF receptor (erbB1) and erbB4 when expressed individually and erbB2 and erbB3 when expressed together. Recombinant NRG-3 altered the growth of human breast cancer cells growing in vitro. NRG-3 was expressed in cell lines derived from breast cancer. These results indicate that NRG-3 is a potential regulator of normal and malignant breast epithelial cells in vivo.


Assuntos
Neoplasias da Mama/metabolismo , Proteínas de Transporte/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular , Proteínas Oncogênicas v-erbB/metabolismo , Sequência de Aminoácidos , Proteínas de Transporte/química , Divisão Celular , Linhagem Celular , Bases de Dados Factuais , Fator de Crescimento Epidérmico/química , Células Epiteliais/metabolismo , Humanos , Dados de Sequência Molecular , Neurregulinas , Proteínas Oncogênicas v-erbB/genética , Conformação Proteica , RNA Mensageiro/metabolismo , Proteínas Recombinantes/metabolismo , Homologia de Sequência de Aminoácidos , Células Tumorais Cultivadas
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