RESUMEN
Some colorectal tumors with wild-type adenomatous polyposis coli gene have activating mutations in beta-catenin (encoded by CTNNB1) that result in decreased phosphorylation by GSK-3beta and increased signaling through the Tcf/Lef transcription factors. To investigate the relationship between CTNNB1 mutations and underlying pathways of genomic instability, we examined 80 colorectal cancers stratified by the presence or absence of microsatellite instability (MSI). CTNNB1 mutations were identified in 13 (25%) of 53 cancers with high frequency MSI (MSI-H), including 12 point mutations at exon 3 phosphorylation sites (codons 41 and 45) and one deletion of the entire exon 3 degradation box. No CTNNB1 mutations were identified in 27 microsatellite stable or low frequency MSI (MSI-L) colorectal cancers (P < 0.01). In contrast, CTNNB1 mutations were identified in 3 of 9 (33%) MSI-H and 10 of 20 (50%) MSS/MSI-L endometrial carcinomas, suggesting a more generalized involvement in these tumors. Only six (46%) of the endometrial carcinoma CTNNB1 mutations occurred at residues directly phosphorylated by GSK-3beta, and only one of these was at either codon 41 or 45. All point mutations in MSI-H cancers were transitions, whereas 64% of those in MSS/MSI-L cancers were transversions (P < 0.01). The differences in the mutation profiles suggest that there may be molecular fingerprints of CTNNB1 mutations, determined by biological factors related to both tumor type and underlying pathways of genomic instability.
Asunto(s)
Adenocarcinoma/genética , Sustitución de Aminoácidos , Transformación Celular Neoplásica/genética , Neoplasias del Colon/genética , Proteínas del Citoesqueleto/genética , Neoplasias Endometriales/genética , Repeticiones de Microsatélite , Mutación Puntual , Transactivadores , Adenoma/genética , Poliposis Adenomatosa del Colon/genética , Diferenciación Celular , Codón/genética , Análisis Mutacional de ADN , ADN de Neoplasias/genética , Exones/genética , Femenino , Genes APC , Humanos , Masculino , Especificidad de Órganos , Fosforilación , Procesamiento Proteico-Postraduccional , Transducción de Señal/genética , beta CateninaRESUMEN
The inability to distinguish recrudescent Plasmodium falciparum infections (treatment failures) from reinfections (new infections) is an important impediment to the evaluation of antimalarial treatment regimens. Ten paired primary and recrudescent isolates collected near the Thai-Cambodian border were analyzed by restriction fragment length polymorphism (RFLP) and by polymerase chain reaction (PCR) genotyping of the genes encoding the following proteins: circumsporozite (CS) protein, erythrocyte binding antigen (EBA)-175, ring-infected erythrocyte surface antigen (RESA), merozoite surface protein-1 (MSP-1), and MSP-2. Both methods demonstrated that the fingerprint pattern of each recrudescent isolate was identical to or was contained within the pattern of the primary isolate. Each recrudescent isolate was unique when compared with the other nine primary isolates. Typing by PCR was more sensitive for the detection of multiclone infections and could be performed with small volumes of whole blood. The PCR genotyping could be a practical method for distinguishing a recrudescent from a new infection when treatment studies are conducted in areas with active malaria transmission.
Asunto(s)
Malaria Falciparum/parasitología , Plasmodium falciparum/genética , Adolescente , Adulto , Animales , Antimaláricos , Dermatoglifia del ADN , Genotipo , Humanos , Malaria Falciparum/tratamiento farmacológico , Masculino , Personal Militar , Plasmodium falciparum/clasificación , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Recurrencia , Insuficiencia del TratamientoRESUMEN
The inability to distinguish failures of treatment of Plasmodium falciparum infection from new infections is an important impediment to the evaluation of antimalarial drugs. On the basis of a pilot study utilizing polymerase chain reaction (PCR) single-strand conformational polymorphism (SSCP) analysis to genotype P. falciparum isolates, we sought to confirm that PCR SSCP analysis could reliably distinguish infections for which treatment failed from unrelated infections with a sample size adequate to estimate the accuracy of this technique. PCR SSCP analysis of the MSP-1, MSP-2, and GLURP genes was performed on 72 paired isolates recovered from 36 individuals for whom treatment failed in Thailand. In every case (100% [95% confidence interval (CI), 90%-100%]), the PCR SSCP pattern of the recrudescent isolates matched that of the primary isolate. We determined whether PCR SSCP analysis could separate unrelated infections by comparing each recrudescent isolate with each of the unrelated primary isolates. Of 1,260 comparisons, 1,258 (99.8% [95% CI, 99.4%-100%]) were unique. The results indicate that PCR SSCP analysis can be used to differentiate infections for which treatment failed from reinfections.