RESUMEN
BACKGROUND: Traditional phenotype-based screening for ß-globin variant and ß-thalassemia using hematological parameters is time-consuming with low-resolution detection. Development of a MALDI-TOF-MS assay using alternative markers is needed. METHODS: We constructed a MALDI-TOF-MS-based approach for identifying various ß-globin disorders and classifying thalassemia major (TM) and thalassemia intermedia (TI) patients using 901 training samples with known HBB/HBA genotypes. We then validated the accuracy of population screening and clinical classification in 2 separate cohorts consisting of 16 172 participants and 201 ß-thalassemia patients. Traditional methods were used as controls. Genetic tests were considered the gold standard for testing positive specimens. RESULTS: We established a prediction model for identifying different forms of ß-globin disorders in a single MALDI-TOF-MS test based on δ- to ß-globin, γ- to α-globin, γ- to ß-globin ratios, and/or the abnormal globin-chain patterns. Our validation study yielded comparable results of clinical specificity (99.89% vs 99.71%), and accuracy (99.78% vs 99.16%) between the new assay and traditional methods but higher clinical sensitivity for the new method (97.52% vs 88.01%). The new assay identified 22 additional abnormal hemoglobins in 69 individuals including 9 novel ones, and accurately screened for 9 carriers of deletional hereditary persistence of fetal hemoglobin or δß-thalassemia. TM and TI were well classified in 178 samples out of 201 ß-thalassemia patients. CONCLUSIONS: MALDI-TOF-MS is a highly accurate, predictive tool that could be suitable for large-scale screening and clinical classification of ß-globin disorders.
Asunto(s)
Hemoglobinas Anormales , Talasemia beta , Humanos , Globinas beta/genética , Talasemia beta/diagnóstico , Talasemia beta/genética , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Hemoglobina Fetal , Hemoglobinas Anormales/análisis , Proteínas PortadorasRESUMEN
MicroRNAs (miRs/miRNAs) play important roles in the occurrence, metastasis and prognosis of multiple types of cancers. However, the specific role of miR302d3p and its underlying mechanism in breast cancer (BC) have not yet been reported. The present study aimed to identify the role of miR302D3p in BC and its potential mechanism using BC cell lines MCF7 and MDAMB231 and normal breast epithelial cell MCF10A. Cancer and paracancerous tissue from patients with BC were also used. Reverse transcriptionquantitative PCR was performed to detect the expression of miR302d3p and transmembrane Bax inhibitor motif containing 6 (TMBIM6). Dualluciferase reporter assays verified the binding sites of miR302d3p and TMBIM6. Immunohistochemistry was used to measure the expression of TMBIM6. Cell transfection techniques were used to overexpress or interfere with miR302d3p and TMBIM6. A Cell Counting Kit8 assay was performed to detect cell viability, and migration was measured using a wound healing assay. Apoptosis was detected by flow cytometry. The expression levels of apoptosisrelated proteins and pathwayrelated proteins were detected by western blotting. The expression of miR302d3p in BC cell lines was found to be downregulated. It was also demonstrated that miR302d3p could inhibit cell viability and migration and promote apoptosis. The expression of TMBIM6 in BC cell lines and tissues was upregulated. Upregulated miR302d3p was shown to inhibit viability and migration, and promote apoptosis by targeting TMBIM6, during which extracellular signalregulated kinase (ERK) and its phosphorylation were inhibited in the ERK signaling pathway in cells. Overall, the present study demonstrated that miR302d3p could regulate the viability, migration and apoptosis of BC cells through regulating TMBIM6mediated ERK signaling pathway.