RESUMEN
Aberrant activation of Wnt/ß-catenin signaling is one of the most frequent abnormalities in human cancer, including breast cancer. The prognostic value of Wnt ligands has never been fully characterized. In this study, we focused on four Wnt ligands, namely Wnt1, Wnt7a, Wnt7b and Wnt9a, which were commonly studied and found pivotal in Wnt/ß-catenin signaling, but seldom explored for their prognostic value. We investigated the expression of Wnt1, Wnt7a, Wnt7b and Wnt9a in breast cancer tissues by using real-time PCR and immunohistochemical analysis, and further identified their prognostic significance. Results demonstrated that only Wnt7b expression level in breast cancer was significantly higher than that of benign breast. Spearman rank-correlation analysis revealed the expression level of Wnt1, Wnt7b and Wnt9a, but not Wnt7a, were all significantly associated with positive lymph nodes. The Kaplan-Meier survival curve demonstrated that patients with high Wnt7b expression had a shorter overall survival (OS) and recurrence-free survival (RFS) than those with low Wnt7b expression. Moreover, the univariate and multivariate analysis revealed that Wnt7b expression was an independent prognostic factor for both OS and RFS of breast cancer patients. In addition, the high expression of Wnt7b in breast cancer and its prognostic role were further validated by GENT (Gene Expression database of Normal and Tumor tissues) database and the Kaplan-Meier plotter database. Taken together, we identified that high expression of Wnt7b, rather than Wnt1, Wnt7a and Wnt9a, may serve as a prognostic biomarker for breast cancer.
RESUMEN
BACKGROUND: Collagen triple helix repeat containing-1 (CTHRC1), which was firstly identified overexpressed in the adventitia and neointima of injured rat arteries, could inhibit collagen expression and increase cell migration. It was then found to be ubiquitously expressed in numerous cell types such as fibroblasts and smooth muscle cells, and aberrantly up-regulated in several malignant tumors. However, the functional role of CTHRC1 and its related mechanism in breast cancer still remains unclear. METHODS: CTHRC1 expressions in breast cancer tissues and cells were assessed by qRT-PCR, western blot and immunohistochemistry. The relative expression level of miR-134, miR-155, miR-30c and miR-630 in breast cancer cells respectively was detected by qRT-PCR. Wild type (Wt) and Mutant type (Mut) CTHRC1 3'UTR sequences were cloned into a psi-CHECK2 reporter vector, and the relative luciferase activity was detected by dual-luciferase reporter assay in indicated cells. The effect of ectopic expression of miR-30c or gain and loss of CTHRC1 on cell viability, cell proliferation, cell cycle progression and apoptosis, cell invasion and migration was respectively detected by CCK-8 assay, colony formation assay, flow cytometry analysis, transwell invasion/migration assay. Protein levels of ß-catenin, active ß-catenin, normal and phosphorylated form of GSK-3ß were detected by western blot in indicated cells. Immunofluorescence staining of ß-catenin was performed to observe nuclear localization. RESULTS: We found CTHRC1 was frequently up-regulated in human breast cancer cells and tissues. Then our cohort study and further meta-analysis validated high expression of CTHRC1 was associated with aggressive clinicopathological features and poor clinical outcome of breast cancer patients. In addition, CTHRC1 promoted cell proliferation, invasion and migration and suppressed cell apoptosis in breast cancer, which might be by activating GSK-3ß/ß-catenin signaling and inhibiting Bax/Caspase-9/Caspase-3 signaling respectively; and these biological functions of CTHRC1 could be directly negatively regulated by miR-30c. CONCLUSION: Taken together, we identified the role of miR-30c/CTHRC1 axis in breast cancer progression and demonstrated CTHRC1 may serve as a prognostic biomarker and therapeutic target for breast cancer.
Asunto(s)
Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Proteínas de la Matriz Extracelular/genética , MicroARNs/genética , Adulto , Anciano , Apoptosis/genética , Biomarcadores , Biomarcadores de Tumor , Neoplasias de la Mama/mortalidad , Neoplasias de la Mama/terapia , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular , Femenino , Humanos , Inmunohistoquímica , Persona de Mediana Edad , Clasificación del Tumor , Metástasis de la Neoplasia , Estadificación de Neoplasias , Oportunidad Relativa , Pronóstico , Interferencia de ARN , Transducción de Señal , Carga TumoralRESUMEN
Glyoxalase 1 (Glo1) is an enzyme that plays a role to metabolize and inactivate methylglyoxal. Previous studies also have confirmed that Glo1 is closely related with tumorigenesis, metastasis, and drug-resistant, but its prognostic value in breast cancer has never been explored. In this study, we investigated the expression of Glo1 in breast cancer cell lines and tissues using real-time PCR, western blot and immunohistochemical analysis. We found Glo1 was frequently up-regulated in human breast cancer cells and tissues, and high expression of Glo1 was associated with positive lymph node, lymphovascular invasion, and TNM stage (all P<0.05). The Kaplan-Meier survival curve demonstrated that patients with high Glo1 expression had a shorter overall survival (OS) and recurrence-free survival (RFS) (Both P<0.001) than those with low Glo1 expression. Moreover, the univariate and further multivariate analysis revealed that Glo1 expression was an independent prognostic factor for both OS and RFS of breast cancer patients. Next, with CCK-8 assay, cell apoptosis analysis, colony formation assay, transwell invasion/migration assay, and wound-healing assay, we validated knock-down of Glo1 suppressed invasion and migration and promoted apoptosis of breast cancer cells. Taken together, we demonstrated the tumor-promoter Glo1 may serve as a prognostic biomarker for breast cancer.
