Cathelicidin-BF regulates the AMPK/SIRT1/NF-κB pathway to ameliorate murine osteoarthritis: In vitro and in vivo studie.

Osteoarthritis (OA) is a chronic degenerative disease with a significant prevalence that causes cartilage damage and can lead to disability. The main factors contributing to the onset and progression of OA include inflammation and degeneration of the extracellular matrix. Cathelicidin-BF (BF-30), a natural peptide derived from Bungarus fasciatus venom, has shown multiple important pharmacological effects. However, the action mechanism of BF-30 in OA treatment remains to be elucidated. In this research, X-ray and Safranin O staining were employed to evaluate the imageology and histomorphology differences in the knee joints of mice in vivo. Techniques such as Western blot analysis, RT-qPCR, ELISA, and immunofluorescence staining were applied to examine gene and protein level changes in in vitro experiments. It was found that BF-30 significantly decreased inflammation and enhanced extracellular matrix metabolism. For the first time, it was demonstrated that the positive effects of BF-30 are mediated through the activation of the AMPK/SIRT1/NF-κB pathway. Moreover, when BF-30 was co-administered with Compound C, an AMPK inhibitor, the therapeutic benefits of BF-30 were reversed in both in vivo and in vitro settings. In conclusion, the findings suggest that BF-30 could be a novel therapeutic agent for OA improvement.

Diagnostic and prognostic potentials of microRNA-27a in osteosarcoma.

AIM: Growing evidence suggest that the microRNA (miR)-23a/24-2/27a cluster may play a crucial role in mammary tumorigenesis and act as a novel class of oncogenes. Among these members, miR-27a has been reported to promote proliferation, migration and invasion in human osteosarcoma cells. The aim of this study was to detect the serum levels of miR-27a in osteosarcoma patients and to investigate its associations with clinicopathological features and prognosis. METHODS: miR-27a levels in sera from 166 osteosarcoma patients and 60 healthy controls were detected by real-time quantitative RT-PCR. Then, the associations of serum miR-27a level with clinicopathological factors or survival of osteosarcoma patients were further evaluated. RESULTS: Compared to healthy controls, the serum levels of miR-27a were significantly increased in osteosarcoma patients (P<0.001). Importantly, miR-27a could efficiently screen osteosarcoma patients from healthy controls (Area under receiver operating characteristic curve, AUC=0.867). Then, high miR-27a expression was more frequently occurred in osteosarcoma patients with advanced clinical stage (P=0.001), positive distant metastasis (P=0.01) and poor response to chemotherapy (P=0.008). In Kaplan-Meier survival analysis, high miR-27a expression was a significant indicator for poor overall survival (P=0.006) as well as poor disease-free survival (P=0.01). Furthermore, multivariate analysis demonstrated that miR-27a expression was an independent and significant prognostic factor to predict overall survival (P=0.01) and disease-free survival (P=0.03). CONCLUSION: miR-27a expression may be elevated in sera of osteosarcoma patients and in turn contributes to aggressive progression of this malignancy. Detection of serum miR-27a levels may have clinical potentials as a non-invasive diagnostic/prognostic biomarker for osteosarcoma patients.

Serum miR-195 is a diagnostic and prognostic marker for osteosarcoma.

BACKGROUND: MicroRNAs have been proved to be successful diagnostic and prognostic markers in several cancers, including osteosarcoma. miR-195, as a tumor suppressor, has been reported to suppress osteosarcoma cell invasion and migration in vitro. However, its clinical value is still unclear. The aim of this study was to determine the expression pattern of miR-195 in osteosarcoma patients and to investigate its associations with tumor progression and prognosis. METHODS: miR-195 levels in sera from 166 osteosarcoma patients and 60 healthy controls were detected by real-time quantitative reverse transcription polymerase chain reaction. Then, the association of serum miR-195 level with clinicopathologic factors or survival of osteosarcoma patients was further evaluated. RESULTS: miR-195 levels in sera from osteosarcoma patients were significantly lower than those in healthy controls (P < 0.001). Importantly, miR-195 could efficiently screen osteosarcoma patients from healthy controls (area under receiver operating characteristic curve = 0.892). Additionally, low miR-195 expression was more frequently occurred in osteosarcoma patients with advanced clinical stage (P = 0.002) and positive distant metastasis (P = 0.008). Univariate survival analysis revealed that osteosarcoma patients with low miR-195 expression had significantly shorter overall survival and disease-free survival than those with high miR-195 expression (both P < 0.001). Multivariate analysis further determined that low miR-195 expression was an independent and significant prognostic factor to predict poor overall survival (P = 0.002) and disease-free survival (P = 0.01). CONCLUSIONS: Decreased expression of miR-195 in serum may be a novel biomarker for screening osteosarcoma and can predict poor prognosis. Detection of serum miR-195 expression may have potential applications for the diagnosis, prognosis, and treatment of osteosarcoma.

Combined microRNA-340 and ROCK1 mRNA profiling predicts tumor progression and prognosis in pediatric osteosarcoma.

To investigate the association of combined microRNA-340 (miR-340) and ROCK1 mRNA profiling with clinicopathologic features and prognosis in pediatric patients with osteosarcoma. Quantitative real-time reverse transcriptase-polymerase chain reaction analysis was performed to detect expression levels of miR-340 and ROCK1 mRNA in cancerous and noncancerous bone tissues from 92 children treated for primary osteosarcomas. Compared with noncancerous bone tissues, the expression levels of miR-340 and ROCK1 mRNA were, respectively, downregulated and upregulated in osteosarcoma tissues (both p < 0.001), which was consistent with the results of in situ hybridization and immunohistochemistry analysis. The downregulation of miR-340 was negatively correlated with the upregulation of ROCK1 mRNA in osteosarcoma tissues (r = -0.78, p = 0.001). In addition, the combined miR-340 downregulation and ROCK1 upregulation (miR-340-low/ROCK1-high) occurred more frequently in osteosarcoma tissues with positive metastasis (p < 0.001) and poor response to pre-operative chemotherapy (p = 0.002). Moreover, miR-340-low/ROCK1-high expression was significantly associated with both shortest overall survival (p < 0.001) and progression-free survival (p < 0.001). Multivariate analysis further confirmed that miR-340-low/ROCK1-high expression was an independent prognostic factor of unfavorable survival in pediatric osteosarcoma (for overall survival: p = 0.006, for progression-free survival: p = 0.008). Our data offer convincing evidence, for the first time, that the combined miR-340 downregulation and ROCK1 upregulation may be linked to tumor progression and adverse prognosis in pediatric osteosarcoma.

Upregulated expression of microRNA-214 is linked to tumor progression and adverse prognosis in pediatric osteosarcoma.

BACKGROUND: MicroRNA-214 (miR-214) expression has been demonstrated to be dysregulated in human malignancies and to play various roles in tumor progression. While previous study of miRNA expression profiling found that it was one of the most upregulated miRNAs in osteosarcoma signature, the potential role of miR-214 in osteosarcomas has been unclear. Therefore, the aim of this study was to investigate association of miR-214 expression with clinicopathologic features and prognosis in pediatric patients with osteosarcoma. PROCEDURE: Quantitative real-time reverse transcriptase-polymerase chain reaction analysis was performed to detect expression levels of miR-214 in cancerous and noncancerous bone tissues from 92 children treated for primary osteosarcomas. Then, the clinical significance of miR-214 dysregulation in pediatric osteosarcomas was also determined. RESULTS: Compared with noncancerous bone tissues, the expression levels of miR-214 were significantly upregulated in osteosarcoma tissues (P < 0.001). High miR-214 expression occurred more frequently in osteosarcoma tissues with large tumor size (P = 0.01), positive metastasis (P = 0.001) and poor response to pre-operative chemotherapy (P = 0.006). Moreover, high miR-214 expression was significantly associated with both shorter overall (P < 0.001) and progression-free survival (PFS; P = 0.001). Multivariate analysis by the Cox proportional hazard model further confirmed that high miR-214 expression was an independent prognostic factor of unfavorable survival in pediatric osteosarcoma (for overall survival: P = 0.008; for PFS: P = 0.01). CONCLUSION: Our data offer evidence that upregulated expression of miR-214 may be linked to tumor progression and adverse prognosis in pediatric osteosarcoma. Further investigation in prospective studies would appear warranted.

Upregulation of SOX9 in osteosarcoma and its association with tumor progression and patients' prognosis.

OBJECTIVE: SOX9 plays an important role in bone formation and tumorigenesis. However, its involvement in osteosarcoma is still unclear. The aim of this study was to investigate the expression pattern and the clinical significance of SOX9 in human osteosarcoma. METHODS: SOX9 mRNA and protein expression levels were detected by RT-PCR and Western blot assays, respectively, using 30 pairs of osteosarcoma and noncancerous bone tissues. Then, immunohistochemistry was performed to analyze the association of SOX9 expression in 166 osteosarcoma tissues with clinicopathological factors or survival of patients. RESULTS: SOX9 expression at mRNA and protein levels were both significantly higher in osteosarcoma tissues than those in corresponding noncancerous bone tissues (both P < 0.001). Immunohistochemical staining indicated that SOX9 localized to the nucleus and high SOX9 expression was observed in 120 of 166 (72.3%) osteosarcoma specimens. In addition, high SOX9 expression was more frequently occurred in osteosarcoma tissues with advanced clinical stage (P = 0.02), positive distant metastasis (P = 0.008) and poor response to chemotherapy (P = 0.02). Osteosarcoma patients with high SOX9 expression had shorter overall survival and disease-free survival (both P < 0.001). Furthermore, the multivariate analysis confirmed that upregulation of SOX9 was an independent and significant prognostic factor to predict poor overall survival and disease-free survival (both P = 0.006). CONCLUSIONS: Our data show for the first time that SOX9 is upregulated in aggressive osteosarcoma tissues indicating that SOX9 may participate in the osteosarcoma progression. More importantly, SOX9 status is a useful prognostic factor for predicting the prognosis of osteosarcoma, suggesting that SOX9 may contribute to the optimization of clinical treatments for osteosarcoma patients. VIRTUAL SLIDES: The virtual slides for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1318085636110837.

MicroRNA-145 downregulation associates with advanced tumor progression and poor prognosis in patients suffering osteosarcoma.

PURPOSE: Microribonucleic acid (miRNA)-145 (miR-145) has been identified as a tumor suppressor in several types of human cancers. Especially, miR-145 expression has been found to be significantly decreased in osteosarcoma tissues, and enforced expression of this miRNA could inhibit invasion and angiopoiesis of osteosarcoma cells. However, its clinical significance in osteosarcoma is still unclear. Therefore, the aim of this study was to analyze the association of miR-145 expression with clinicopathologic features and prognosis in patients suffering osteosarcoma. METHODS: miR-145 expression was detected by quantitative real-time reverse transcriptase polymerase chain reaction analysis using 166 pairs of osteosarcoma and noncancerous bone tissues. Then, the associations of miR-145 expression with clinicopathological factors or survival of patients suffering osteosarcoma were determined. RESULTS: The expression levels of miR-145 in osteosarcoma tissues were significantly lower than those in corresponding noncancerous bone tissues (P < 0.0001). In addition, miR-145 downregulation more frequently occurred in osteosarcoma specimens with advanced clinical stage (P = 0.003) and positive distant metastasis (P = 0.008). Moreover, the univariate analysis demonstrated that osteosarcoma patients with low miR-145 expression had poorer overall (P = 0.003) and disease-free survival (P < 0.001). Furthermore, the multivariate analysis identified low miR-145 expression as an independent prognostic factor for both overall (P = 0.01) and disease-free survival (P = 0.008). CONCLUSION: For the first time, the current data offer convincing evidence that the down-regulation of miR-145 may be associated with tumor aggressiveness and tumor metastasis of osteosarcoma, and that this miRNA may be an independent prognostic marker for osteosarcoma patients.

Loss of microRNA-132 predicts poor prognosis in patients with primary osteosarcoma.

MicroRNA-132 (miR-132), an angiogenic growth factor inducible microRNA in the endothelium, facilitates pathological angiogenesis. Previous study showed that miR-132 was downregulated in human osteosarcoma. However, its functional attributes associated with tumor progression of osteosarcoma have not been fully elucidated. The aim of this study was to investigate the clinical significance of miR-132 expression in human osteosarcoma. miR-132 expression was detected by quantitative reverse transcription polymerase chain reaction using 166 pairs of osteosarcoma and noncancerous bone tissues. Then, the association of miR-132 expression with clinicopathological factors or survival of osteosarcoma patients was also evaluated. miR-132 expression was significantly lower in osteosarcoma tissues than that in corresponding noncancerous bone tissues (P < 0.001). In addition, miR-132 expression was decreased in the osteosarcoma specimens with advanced clinical stage (P = 0.009), positive distant metastasis (P = 0.006), and poor response to chemotherapy (P = 0.009). Moreover, both the univariate and multivariate analyses showed that osteosarcoma patients with low miR-132 expression had poorer overall and disease-free survival (both P < 0.001), and low miR-132 expression was an independent prognostic factor for both overall (P = 0.001) and disease-free survival (P = 0.006). These findings offer the convinced evidence for the first time that miR-132 may participate in tumor progression of osteosarcoma and loss of miR-132 expression may be a predictor for unfavorable outcome of osteosarcoma patients.

Prognostic evaluation of microRNA-210 expression in pediatric osteosarcoma.

MicroRNA-210 (miR-210) plays important roles in the regulation of cell growth, angiogenesis, and apoptosis in different cancer type. Previous study of miRNA expression profiling found that miR-210 was significantly elevated in osteosarcoma samples. However, its roles in this disease have not been fully elucidated. Thus, the aim of this study was to investigate the association of miR-210 expression with clinicopathologic features and prognosis in patients with osteosarcomas. Quantitative real-time reverse transcriptase-polymerase chain reaction analysis was performed to detect the expression level of miR-210 in cancerous and noncancerous bone tissues from 92 children treated for primary osteosarcomas. MiR-210 expression was significantly increased in osteosarcoma tissues compared with that in corresponding noncancerous bone tissues (P < 0.001). In addition, miR-210 upregulation more frequently occurred in osteosarcoma tissues with large tumor size (P = 0.02), poor response to preoperative chemotherapy (P = 0.008), and positive metastasis (P = 0.01). Moreover, miR-210 upregulation was associated with significantly decreased overall survival (P = 0.007) and progression-free survival (P = 0.01). In the Cox proportional hazard model, it was confirmed that its expression in the biopsy samples was an independent prognostic factor of unfavorable survival in osteosarcoma (for overall survival: P = 0.01; for progression-free survival: P = 0.02). These findings suggested that miR-210 upregulation showed a strong correlation with tumor aggressive progression of pediatric osteosarcoma and could help prognostic screening of patients with this malignancy.