Browsing by Subject "Osteosarcoma"

Now showing 1 - 7 of 7
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    An association between successful engraftment of osteosarcoma patient-derived xenografts and clinicopathological findings
    (Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Fortuna-Costa, Anneliese; Alcantara Granato, Regina; Meohas, Walter; de Sá Lopes, Ana Cristina; Cunha Caruso, Anabela; Castro e Silva Pinheiro, Rafael; da Gama d'Eça, Pedro; Braga Dias, Rhayra; Perini, Jamila Alessandra; Fernandes Barbosa, Ana Paula; Moreira de Sá, Renato Augusto; Matheus Guimarães, João Antonio; Murray, Samuel S.; Leite Duarte, Maria Eugenia
    Although osteosarcoma is a rare disease, with a global incidence rate estimated at 5.0/million/ year, it is the most frequent primary bone sarcoma in children and adolescents. In translational research, the patient-derived xenograft (PDX) model is considered an authentic in vivo model for several types of cancer, as tumorgrafts faithfully retain the biological characteristics of the primary tumors. Our goal was to investigate the association between PDX formation and clinical findings of osteosarcoma patients and the ability of the model to preserve in immunocompromised mice the characteristics of the parental tumor. A fresh sample of the patient tumor obtained from a representative biopsy or from surgical resection was implanted into nude mice. When tumor outgrowths reached ~1,500 mm 3 , fresh PDX fragments were re-transplanted into new hosts. Engraftment in mice was obtained after a latency period of 19-225 days (median 92 days) in 40.54% of the implanted samples. We confirmed the histopathological fidelity between the patient tumor and their respective established PDXs, including the expression of biomarkers. PDX take rate was higher in surgical resection samples, in post-chemotherapy surgical samples and in samples from patients with metastatic disease at presentation. In conclusion, we have shown that the osteosarcoma PDX model reliably recapitulates the morphological aspects of the human disease after serial passage in mice. The observation that more aggressive forms of osteosarcoma, including those with metastatic disease at presentation, have a higher efficiency to generate PDXs provides a promising scenario to address several unanswered issues in clinical oncology.
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    Diallyl trisulfide regulates cell apoptosis and invasion in human osteosarcoma U2OS cells through regulating PI3K/AKT/GSK3β signaling pathway
    (Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) He, Pan; Wang, Zhijun; Sheng, Bin; Xu, Yongqiang; Feng, Siyin; Huang, Yan; Gong, Fuqiang; Tang, Liting; Xie, Liming
    Aims. To investigate the effects and the mechanisms of action of Diallyl trisulfide (DATS) on the proliferation and metastasis of human osteosarcoma (OS) U2OS. Methods. U2OS cells were treated by different concentrations of DATS at different time points. Cell proliferations were measured by MTT assay. DATS induced cell cycle distribution and apoptosis were evaluated by flow cytometry (FCM) with Annexin-V. Cell migration and invasion were detected by wound healing assay and transwell assay. The effects of DATS in U2OS cell growth and metastasis were also detected in a mouse OS xenograft model. Results. A time- and concentration-dependent cytotoxic effect of DATS was observed in U2OS cells. FCM with PI staining and Annexin-V -FITC indicated that DATS induces apoptosis and a G0/G1 cell cycle arrest of U2OS cells at all concentrations from 25 μmol/l to 100 μmol/l. DATS also inhibits the migration and invasion of U2OS cells. Western blot showed that the expression levels of p-AKT, p-GSK3β, Bcl-2, Vimentin and β-catenin were decreased, while the expression levels of Bad, Bax and E-cadherin were significantly increased in DATS treated U2OS cells. Analysis using a mouse xenograft model indicated that xenografts of DATS treatment group had a significant decrease in tumor volume and weight compared to the control group. Lung metastasis models in mice demonstrated that treatment of DATS inhibits lung metastasis of OS in vivo. Conclusions. These data suggested that DATS inhibits OS development and progression through the regulation of PI3K/AKT/GSK3β signaling pathways, accompanied by downregulation of Bcl-2, Vimentin and β-catenin, as well as upregulation of Bad, Bax and Ecadherin. Therefore, our data demonstrated that DATS exerted its anticancer effects by inhibiting cell proliferation, migration and invasion in vitro and in vivo. These results provide evidence for the use of the natural product DATS either alone or in combination with standard therapy for OS.
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    Expression of CXCR4 and MMP-2 is associated with poor prognosis in patients with osteosarcoma
    (Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Gong, Chen; Sun, Kai; Xiong, Hui-hua; Sneh, Tal; Zhang, Jing; Zhou, Xiao; Yan, Peng; Wang, Jian-hua
    Backgroud. Osteosarcoma is a primary malignant tumor with a high tendency to form metastasis and poor prognosis. Consequently, finding effective early indicators of metastases is crucial for identifying and treating high-risk patients. CXCR4 and MMP-2 have been found to strongly correlate with invasion and metastasis of malignant tumors, including osteosarcoma. Materials and Methods. Our study evaluated CXCR4 in conjunction with MMP-2 as an important clinicopathological prognostic predictor for metastasis and overall survival of osteosarcoma. 73 patients’ clinical data and pathological samples were retrieved for the study. A median time of 36 months follow-up was performed to evaluate for tumor metastasis and patient survival. CXCR4 and MMP-2 proteins in tumor tissues were detected by immunohistochemistry on paraffin- embedded tissue sections. Results. The positive expression rate of CXCR4 and MMP-2 was 68.5% and 54.8% respectively, and of the 45 patients who developed distal metastasis, 33 and 28 patients had positive expression of CXCR4 and MMP-2 respectively. The median metastasis-free survival was 72.00 months in the CXCR4-negative group and 14.00 months in the CXCR4 positive group. Furthermore, median overall survival was 73.77 and 24.00 months in these same two groups. Further, the median metastasis-free survival was 66.51 months in the MMP-2 negative group and 9.00 months in the MMP-2 positive group. The median overall survival was 75.07 and 19.00 months in these same two groups. MMP2 and metastasis remained the significant and independent prognostic factors for metastasis-free survival and overall survival by using the COX regression model adjusted for the multivariate predictors of survival. Conclusion. Our results suggest that metastasis and MMP-2 are both independent prognostic indicators for metastasis-free and overall survival of osteosarcoma patients.
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    Ezrin and moesin expression in canine and feline osteosarcoma
    (Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Hlavaty, Juraj; Wolfesberger, Birgitt; Hauck, Marlene; Obermayer Pietsch, Barbara; Fuchs Baumgartinger, Andrea; Miller, Ingrid; Walter, Ingrid
    Biological features of canine osteosarcomas (OS) differ markedly from those found in feline and resemble more human osteosarcomas, in particular for their high rate of metastasis and poor prognosis. Ezrin, radixin and moesin are members of the ERM protein family and link the actin cytoskeleton with the cell membrane. Ezrin and moesin have been shown to be of prognostic significance in tumor progression due to their role in the metastatic process. The objective of this study was to analyze ezrin and moesin protein expression in a series of dog (n=16) and cat (n=8) osteosarcoma samples using immunohistochemistry and western blot techniques. We found that cat OS have a higher moesin expression compared to dog OS, however, the active phosphorylated forms of moesin and ezrin Tyr353 were more abundant in the dog samples. A statistically significant difference was found for the low and high immunohistochemical scores of ezrin and pan-phosphoERM proteins between cat and dog. Although phosphoezrin Thr567 was higher in feline OS, the membranous localization in dog OS samples indicates the presence of the biologically active form. Therefore, the observed differences in phosphorylated forms of ezrin and moesin status should be further studied to demonstrate if they are relevant for different biological behavior between dog and cat OS.
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    Methylation of histone H3 lysine 27 associated with apoptosis in osteosarcoma cells induced by staurosporine
    (Murcia : F. Hernández, 2009) Cheng, Ming-Fang; Lee, Chian-Her; Hsia, Kan-Tai; Huang, , Guo-Shu; Lee, Herng-Sheng
    The relationship between histone methylation and apoptosis, programmed cell death, is beginning to be explored. The objective of this study was to investigate the effects of staurosporine, a PKC inhibitor on the methylation of histone H3 in osteosarcoma cells. Following stimulation by staurosporine in vitro of G292 cells, a human osteosarcoma cell line with fibroblast-like phenotype, methylation of histone H3 was evaluated by western blotting and immunocytochemistry. G292 cells revealed the expression of cleaved PARP after incubation with staurosporine for 3 hours. Monomethyl lysine (K) 27 was induced by staurosporine at a concentration of 1, but no monomethyl K4 or K9 in histone H3 was seen. Dimethyl and trimethyl histone H3 K27 were also identified. There was no expression of dimethyl or trimethyl histone H3 K4 and K9. Expression of monomethyl histone H3 K27 was dose-dependent. The morphologic changes of apoptosis induced by staurosporine were observed under microscopy. Immunocytochemistry of monomethyl histone H3 K27 showed a weak signal in controls, a strong signal in staurosporine-treated tumor cells and a denser signal in the apoptotic cells. Our studies demonstrated that monomethyl histone H3 lysine 27 is expressed in staurosporine-induced apoptotic osteosarcoma cells. The findings may provide novel bridge information between the epigenetic episodes and apoptotic process
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    Silencing of synaptotagmin 7 regulates osteosarcoma cell proliferation, apoptosis, and migration
    (Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Wu, Zhiqiang; Sun, Zhengwang; Huang, Rui; Zang, Ding; Wang, Chunmeng; Yan, Xu; Yan, Wangjun
    Background. Synaptotagmin 7 (SYT7) is a component of the synaptotagmin family, which is essential in many physiological and pathological processes. In this study, we aimed to investigate the role of SYT7 in osteosarcoma. Methods. We defined the expression levels of SYT7 in osteosarcoma tissues and para-sarcoma tissues by immunohistochemistry and analyzed the possible correlation between SYT7 expression and pathological characteristics via Mann-Whitney U analysis and Spearman correlation analysis. The effects of SYT7 silencing in vitro on cell growth were assessed by MTT assay. Cell cycle and cell apoptosis were assessed by flow cytometry analysis. Wound healing assay and transwell assay were applied to assess the migration and invasion capacity. Results. The results showed that the expression levels of SYT7 were upregulated in osteosarcoma tissues compared with para-sarcoma tissues and positively correlated with the pathological characteristics of osteosarcoma. Functional experiments demonstrated that SYT7 silencing significantly inhibited cell proliferation and colony formation capacity (P<0.001), induced cell cycle arrest which increased the proportion of G2 phase and decreased the proportion of S phase, enhanced cell apoptosis (P<0.01), and limited the capacity of migration and invasion (P<0.01), compared with shCtrl group. Conclusion. The results indicated that SYT7 plays a crucial role in the development of osteosarcoma. SYT7 can be applied as a new diagnostic and therapeutic target in osteosarcoma.
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    The expression of hydrogen sulfide-producing enzymes in primary and lung metastatic osteosarcoma
    (Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2022) Gong, Lihua; Sun, Xiaoqi; Zhang, Ming; Du, Junbao; Ding, Yi; Jin, Hongfang
    Background. Hydrogen sulfide (H2S) is a novel gas transmitter signaling molecule. H2S is synthesized by cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE), and 3-mercaptopyruvate sulfurtransferase (MST). There have been no reports about the roles of these enzymes in osteosarcoma and its metastases. We detected H2S synthase expression levels in human primary osteosarcoma and lung metastatic osteosarcoma. Methods. Immunohistochemistry was performed in primary osteosarcoma (n=19), lung metastatic osteosarcoma (n=11), osteoblastoma (n=10) and bony callus (n=2). The expression of CBS, CSE, and MST was defined as negative, moderately positive and strongly positive. Results. MST staining was moderately to strongly positive in all cases. CSE staining was negative in 94.7% (18/19) of primary osteosarcoma cases and 90.9% (10/11) of lung metastatic osteosarcoma cases. CBS staining was strongly positive in 68.4% (13/19) of primary osteosarcoma cases, moderately positive in 15.8% (3/19) of cases, and negative in 15.8% (3/19) of cases. In lung metastatic osteosarcoma, the proportions of negative, moderately positive and strongly positive cases were 63.6% (7/11), 18.2% (2/11) and 18.2% (2/11), respectively. Conclusions. CBS and CSE expression, especially CSE expression, decreased in both primary osteosarcoma and lung metastatic osteosarcoma, which may suggest that CBS and CSE play roles in osteoblast cell malignant transformation and osteosarcoma progression. These enzymes could be used as new prognostic assessment factors and may represent new therapeutic targets for osteosarcoma and metastasis prevention