RESUMO
A replication-deficient adenovirus encoding human interferon alpha2b, driven by the human cytomegalovirus (CMV) promoter, was constructed and characterized. This construct was used to infect human cells derived from different types of cancer. The production of protein and its secretion into the culture medium were tested by Western blotting and immunoassay. Inhibition of cell proliferation and antiviral activity, two of the most important biological activities of interferon, were observed with this construct. PC-3 cells, derived from human prostatic cancer, or Hep3B cells, derived from human hepatocellular carcinoma, were injected subcutaneously to generate and establish in vivo tumors in athymic nude mice. Intratumoral injection with the recombinant adenovirus expressing interferon alpha2b resulted in complete regression of tumor growth. Our results demonstrate that interferon gene delivery using recombinant adenoviral vectors may be a useful approach to treat a variety of cancers.
Assuntos
Terapia Genética/métodos , Interferon-alfa/genética , Adenoviridae/genética , Animais , Western Blotting , Divisão Celular , Citomegalovirus/genética , Feminino , Vetores Genéticos , Humanos , Imunoensaio , Interferon alfa-2 , Interferon-alfa/biossíntese , Camundongos , Camundongos Endogâmicos BALB C , Neoplasias Experimentais/tratamento farmacológico , Proteínas Recombinantes , Fatores de Tempo , Células Tumorais CultivadasRESUMO
A novel human cDNA encoding a putative nucleotide-binding protein (NBP) was obtained by screening a human SHSY5Y neuroblastoma library. The deduced protein contains 320 amino acids (aa) with a M(r) of 34,540. NBP displays sequence similarity with the product of the minD gene from Escherichia coli. MinD is involved in the proper placement of the division septum, and has ATPase activity. NBP and MinD contain consensus nucleotide (nt)-binding domains. The NBP mRNA is approx. 1500 nt in length and is expressed in several human cell lines and in all rat tissues examined, with the highest levels in lung and testis.
Assuntos
Adenosina Trifosfatases/genética , Proteínas de Transporte/genética , Proteínas de Escherichia coli , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Sequência Consenso , DNA Complementar , Escherichia coli , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Dados de Sequência Molecular , Nucleotídeos/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Células Tumorais CultivadasRESUMO
A cDNA corresponding to human ribosomal protein L37a (hL37a) was obtained by screening a SHSY5Y neuroblastoma library. The full-length cDNA contained 366 nucleotides (nt) in addition to a poly(A) tail. The pyrimidine-rich sequence, CTTTCT, that is common to many ribosomal protein-encoding cDNAs was present at the 5' terminus. The nt sequence displayed 85% identity with rat L37a (rL37a) cDNA. The predicted protein contains 92 amino acids with a M(r) of 10,277, is highly basic, and has 100% sequence identity with rL37a. A putative zinc-finger domain is present in the central region of the protein. Human lymphocytes and several human cell lines express hL37a mRNA at significantly higher levels than the rat cell lines and rat tissues tested. The hL37a gene does not contain introns.
