RESUMEN
OBJECTIVE: The aim of this study was to investigate the effect of protocadherin 10 (PCDH10) on the invasive potential of lymphoma cells by regulating matrix metalloproteinase-7 (MMP7) and MMP9 via targeting ß-catenin. MATERIALS AND METHODS: The mRNA and protein expressions of PCDH10, ß-catenin, MMP7 and MMP9 in lymphoma cell lines were examined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot, respectively. Raji cells with low expression of PCDH10 were transiently transfected with pCMV5-HA-PCDH10 or pcDNA5-His-ß-catenin. Meanwhile, HUT-78 cells with high expression of PCDH10 were transfected with PCDH10-shRNA or ß-catenin-shRNA. Subsequently, the expression levels of ß-catenin, MMP7 and MMP9 in transfected lymphoma cells were determined as well. In addition, the regulatory effects of PCDH10 on the invasive potential of lymphoma cells were explored by transwell assay. RESULTS: PCDH10 expression was negatively correlated with the expressions of ß-catenin, MMP7 and MMP9 in several lymphoma cell lines. Transfection of HA-PCDH10 in human-derived malignant B lymphoma cell line Raji markedly down-regulated the protein levels of ß-catenin, MMP7 and MMP9. However, the mRNA level of ß-catenin was not influenced by PCDH10. Interference with PCDH10 in HUT-78 cells with high expression of PCDH10 significantly increased the protein expressions of ß-catenin, MMP7, and MMP9. However, no significant changes were observed in the mRNA expression of ß-catenin. In addition, knockdown of ß-catenin in cells with high expression of PCDH10 remarkably down-regulated the expression levels of MMP7 and MMP9. CONCLUSIONS: PCDH10 overexpression in lymphoma cells downregulates ß-catenin expression, as well as inhibits the expressions of MMP7 and MMP9, eventually inhibiting the invasive potential of lymphoma cells.
Asunto(s)
Cadherinas/metabolismo , Regulación hacia Abajo , Linfoma/metabolismo , beta Catenina/metabolismo , Cadherinas/genética , Perfilación de la Expresión Génica , Humanos , Linfoma/patología , Metaloproteinasa 7 de la Matriz/genética , Metaloproteinasa 7 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/metabolismo , Protocadherinas , ARN Mensajero/genética , ARN Mensajero/metabolismo , Células Tumorales CultivadasRESUMEN
Phenotypic variations in α-thalassemia mainly depend on the defective α-globin gene number. Genetic modifiers of the phenotype of Hemoglobin H (HbH) disease were poorly reported, apart from ß-thalassemia allele that was identified ameliorating the severity of α-thalassemia. Because erythroid Krüppel-like factor (KLF1) mutations can modulate the red blood phenotype, we evaluated its effect on the α-thalassemia phenotype. Overall, we identified 72 subjects with five different KLF1 heterozygous mutations in 1468 individuals, including 65 out of 432 α-thalassemia carriers with fetal hemoglobin (HbF) levels ≥1%, 0 out of 310 carriers with HbF levels <1% and 7 out of 726 HbH disease patients. We firstly established the link between KLF1 mutations and relatively elevated hemoglobin A2 (HbA2 ) and HbF levels, along with lower mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) values in a group of α-thalassemia carriers. However, we concluded that KLF1 mutations were not significantly linked to HbH disease severity. On the basis of HBA or HBB genotype and gender, clinical severity of patients with HbH disease was correctly predicted in 73.3% cases. It may improve the screening and diagnostic assessment of α-thalassemia.