RESUMEN
The evolution of gastric carcinogenesis remains largely unknown. We established two gastric carcinogenesis models in New-World nonhuman primates. In the first model, ACP03 gastric cancer cell line was inoculated in 18 animals. In the second model, we treated 6 animals with N-methyl-nitrosourea (MNU). Animals with gastric cancer were also treated with Canova immunomodulator. Clinical, hematologic, and biochemical, including C-reactive protein, folic acid, and homocysteine, analyses were performed in this study. MYC expression and copy number was also evaluated. We observed that all animals inoculated with ACP03 developed gastric cancer on the 9(th) day though on the 14(th) day presented total tumor remission. In the second model, all animals developed pre-neoplastic lesions and five died of drug intoxication before the development of cancer. The last surviving MNU-treated animal developed intestinal-type gastric adenocarcinoma observed by endoscopy on the 940(th) day. The level of C-reactive protein level and homocysteine concentration increased while the level of folic acid decreased with the presence of tumors in ACP03-inoculated animals and MNU treatment. ACP03 inoculation also led to anemia and leukocytosis. The hematologic and biochemical results corroborate those observed in patients with gastric cancer, supporting that our in vivo models are potentially useful to study this neoplasia. In cell line inoculated animals, we detected MYC immunoreactivity, mRNA overexpression, and amplification, as previously observed in vitro. In MNU-treated animals, mRNA expression and MYC copy number increased during the sequential steps of intestinal-type gastric carcinogenesis and immunoreactivity was only observed in intestinal metaplasia and gastric cancer. Thus, MYC deregulation supports the gastric carcinogenesis process. Canova immunomodulator restored several hematologic measurements and therefore, can be applied during/after chemotherapy to increase the tolerability and duration of anticancer treatments.
Asunto(s)
Cebus , Modelos Animales de Enfermedad , Neoplasias Experimentales/patología , Neoplasias Gástricas/patología , Animales , Línea Celular , Venenos de Crotálidos/administración & dosificación , Venenos de Crotálidos/farmacología , Dosificación de Gen/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Inmunohistoquímica , Hibridación Fluorescente in Situ , Metilnitrosourea , Neoplasias Experimentales/diagnóstico por imagen , Neoplasias Experimentales/genética , Extractos Vegetales/administración & dosificación , Extractos Vegetales/farmacología , Proteínas Proto-Oncogénicas c-myc/genética , Proteínas Proto-Oncogénicas c-myc/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Neoplasias Gástricas/diagnóstico por imagen , Neoplasias Gástricas/genética , UltrasonografíaRESUMEN
1-Deoxy-d-xylulose 5-phosphate reductoisomerase (DXR, EC: 1.1.1.267) is the second enzyme in the 2C-methyl-d-erythritol 4-phosphate (MEP) pathway, one of the two pathways in plants that can produce isoprenoids. The MEP pathway is the source of isoprene emitted from leaves, but rubber production is believed to result primarily from the mevalonic acid (MVA) pathway. Two cDNAs for DXR designated HbDXR1 and HbDXR2 were isolated from leaves and latex of rubber tree using RT-PCR based methods. Both cDNAs contain an open reading frame (ORF) of 1416bp encoding 471 amino acids with a molecular mass of about 51kDa. The deduced HbDXRs show extensive sequence similarities to that of other plant DXRs (73-87% identity). Molecular modeling revealed that the two HbDXRs contain all typical characteristics of DXR and share spatial structures, which are very similar to that of Escherichia coli DXR. Phylogenetic and DNA gel blot analyses suggested that a duplication of the DXR gene has occurred in the rubber tree. Semi-quantitative RT-PCR analysis showed that the HbDXR genes are differentially regulated in various tissues of the rubber tree. The HbDXR2 was more highly expressed in clone RRIM 600 than in the wild type, and this is consistent with higher rubber content of this clone. While 2-chloroethane phosphonic acid (ethephon) significantly increased latex yield, it only transiently induced the HbDXR2 gene. The expression of HbDXR2 in the latex suggests its important role in isoprenoid biosynthesis by substrate molecules, indicating that the MEP pathway may have some indirect roles in the biosynthesis of rubber.