Bone transport using the Ilizarov method for osteosarcoma patients with tumor resection and neoadjuvant chemotherapy.

BACKGROUND: Studies on the applications of bone transport using the Ilizarov method for osteosarcoma (OS) patients with surgical resection and neoadjuvant chemotherapy are rare. METHODS: A retrospective analysis was conducted in 10 patients with limb OS receiving limb-salvage treatment by Ilizarov method from 2007 to 2012 in our hospital. The general information, treatment outcomes and follow-up data of the patients were collected. RESULTS: The mean length of the transported fragment and the mean transport distance of the affected limb were both 14 cm. The mean time in the external fixator was 34.2 ±â€¯11.2 months (16-47 months) and the mean external fixation index (EFI) was 75 days/cm. The mean follow-up time was 68.6 ±â€¯26.6 months (37-103 months). Seven patients underwent additional operations to treat the postoperative complications, and the mean number of operation was 1.7 times. Only one patient underwent amputation due to tumor relapse and all patients survived without tumor. The limb-salvage rate was 90%. At the time of external fixator removal, the ASAMI-bone score was good in 66.7% of patients and the ASAMI-function score was fair in 66.7% of cases. The mean MSTS score was 18.6 ±â€¯3.2 (n = 9). At 10 months after fixator removal, both the ASAMI-bone score and ASAMI-function score were both excellent in 80% and good in 20% cases, and the mean MSTS score was further improved to 27.2 ±â€¯1.11 (n = 5). CONCLUSION: Bone transport using the Ilizarov method can achieve good therapeutic effectiveness in the limb-salvage treatment for OS patients with neoadjuvant chemotherapy as long as the complications can be timely recognized and well managed.

Hippo/YAP signaling pathway is involved in osteosarcoma chemoresistance.

BACKGROUND: Osteosarcoma is the most common bone malignancy in children and adolescents, and 20%-30% of the patients suffer from poor prognosis because of individual chemoresistance. The Hippo/yes-associated protein (YAP) signaling pathway has been shown to play a role in tumor chemoresistance, but no previous report has focused on its involvement in osteosarcoma chemoresistance. This study aimed to investigate the role of the Hippo/YAP signaling pathway in osteosarcoma chemoresistance and to determine potential treatment targets. METHODS: Using the Cell Titer-Glo Luminescent cell viability assay and flow cytometry analysis, we determined the proliferation and chemosensitivity of YAP-overexpressing and YAP-knockdown osteosarcoma cells. In addition, using western blotting and the real-time polymerase chain reaction technique, we investigated the alteration of the Hippo/YAP signaling pathway in osteosarcoma cells treated with chemotherapeutic agents. RESULTS: Mammalian sterile 20-like kinase 1 (MST1) degradation was increased, and large tumor suppressor kinase 1/2 (LATS1/2) total protein levels were decreased by methotrexate and doxorubicin, which increased activation and nuclear translocation of YAP. Moreover, YAP increased the proliferation and chemoresistance of MG63 cells. CONCLUSIONS: The Hippo/YAP signaling pathway plays a role in osteosarcoma chemoresistance, and YAP is a potential target for reducing chemoresistance.

Amputation Versus Limb-Salvage Surgery in Patients with Osteosarcoma: A Meta-analysis.

BACKGROUND: This meta-analysis compared survival and function in patients with limb osteosarcoma treated with limb-salvage surgery (LSS) versus amputation or rotationplasty. METHODS: Medline, Cochrane, EMBASE, and Google Scholar were searched until November 30, 2015 for studies reporting Musculoskeletal Tumor Society (MSTS) scores and survival rates in osteosarcoma patients. Differences between patients undergoing LSS versus ablative surgery were analyzed based on MSTS scores and postoperative survival rates. RESULTS: Of 1330 patients in the studies analyzed, 934 underwent LSS, and 662 were treated with amputation. A random-effects model was applied due to heterogeneity among studies (Q statistic = 1.829, I (2) = 0 %, p = 0.767). No difference was found in post-operative local recurrence rate between amputees and patients receiving LSS. The 5-year survival rate was significantly lower with amputation compared with LSS (OR 0.628; 95 % CI 0.431-0.913, p = 0.015). The 2-year survival rate was not different between amputation and LSS. In addition, amputees had lower MSTS scores than those undergoing LSS (difference in means = -4.46 %, 95 % CI 6.49-2.45 %, p < 0.001). CONCLUSIONS: LSS results in higher 5-year survival rates and better functional outcomes as indicated by MSTS scores in patients with limb osteosarcomas.

A high risk of osteosarcoma in individuals who are homozygous for the p.D104N in endostatin.

The D104N polymorphism (p.D104N) in endostatin has been previously identified in many types of cancer, and this polymorphism is believed to be a phenotypic modulator in some tumors. However, it is unknown whether endostatin p.D104N affects the risk and progression of osteosarcoma (OS). Here, we analyzed the p.D104N endostatin variant in 236 patients with OS and 418 healthy individuals. Similar frequencies of wild type and heterozygous p.104DN endostatin were observed in controls and OS patients. Interestingly, the frequency of the homozygous p.D104N (p.104NN) genotype was higher in OS patients group compared to control group, suggesting that individuals with p.104NN endostatin have a significantly increased risk for OS. In addition, OS patients with p.104NN endostatin had a shorter survival time and a higher rate of metastasis than OS patients with wild type endostatin. Animal experiments revealed that overexpression of p.104NN endostatin did not significantly inhibit OS lung metastasis. Interestingly, administration of endostatin dramatically inhibited OS lung metastasis in the p.104NN endostatin xenograft model. Together, these results suggest that p.104NN of endostatin is associated with the risk of OS and demonstrates predictive significance for clinical outcome in OS patients. In addition, endostatin therapy may be necessary for OS patients harboring p.104NN endostatin.

Development of the affinity materials for phosphorylated proteins/peptides enrichment in phosphoproteomics analysis.

Reversible protein phosphorylation is a key event in numerous biological processes. Mass spectrometry (MS) is the most powerful analysis tool in modern phosphoproteomics. However, the direct MS analysis of phosphorylated proteins/peptides is still a big challenge because of the low abundance and insufficient ionization of phosphorylated proteins/peptides as well as the suppression effects of nontargets. Enrichment of phosphorylated proteins/peptides by affinity materials from complex biosamples is the most widely used strategy to enhance the MS detection. The demand of efficiently enriching phosphorylated proteins/peptides has spawned diverse affinity materials based on different enrichment principles (e.g., electronic attraction, chelating). In this review, we summarize the recent development of various affinity materials for phosphorylated proteins/peptides enrichment. We will highlight the design and fabrication of these affinity materials, discuss the enrichment mechanisms involved in different affinity materials, and suggest the future challenges and research directions in this field.

miR-382 inhibits osteosarcoma metastasis and relapse by targeting Y box-binding protein 1.

Lung metastasis and relapse in osteosarcoma (OS) patients indicate poor prognosis. Here, we identified significantly decreased expression of miR-382 in highly metastatic OS cell lines and relapsed OS samples compared to their parental cell lines and primary OS samples, respectively. In addition, our clinical data showed that the miR-382 expression level was inversely associated with relapse and positively associated with metastasis-free survival in OS patients. The overexpression of miR-382 suppressed epithelial-mesenchymal transition (EMT) and metastasis. This overexpression also decreased the cancer stem cell (CSC) population and function in OS cells. In contrast, inhibition of miR-382 stimulated EMT and metastasis and increased CSC population in OS cells. In addition, our in vivo experiments showed that the overexpression of miR-382 inhibited CSC-induced tumor formation, and the combination of miR-382 with doxorubicin prevented disease relapse in OS patients. Furthermore, we demonstrated that miR-382 exerted its tumor-suppressing potential by directly targeting Y box-binding protein 1 (YB-1) in OS. Taken together, our findings suggest that miR-382 functions as a tumor suppressor function and that the overexpression of miR-382 is a novel strategy to inhibit tumor metastasis and prevent CSC-induced relapse in OS.

miR-382 inhibits tumor growth and enhance chemosensitivity in osteosarcoma.

Dysregulation of miRNAs is involved in osteosarcoma (OS). Here, we demonstrate that miR-382 is decreased in specimens of OS patients with a poor chemoresponse compared to those with a good chemoresponse. In addition, our clinical data show that decreased miR-382 was associated with poor survival in OS patients. Overexpression of miR-382 inhibited cell growth and chemoresistance by targeting KLF12 and HIPK3, respectively. In contrast, inhibition of miR-382 or overexpression of target genes stimulated OS cell growth and chemoresistance both in vitro and in vivo. Taken together, these findings suggest that miR-382 is a tumor suppressor miRNA and induction of miR-382 is a potential strategy to inhibit OS progression.

MiR-34c inhibits osteosarcoma metastasis and chemoresistance.

Studies have shown that miR-34c is associated with metastasis and the chemoresponse of several cancers, but its role in osteosarcoma (OS) is unclear. Here, we investigated the role and mechanism of miR-34c in OS metastasis and chemoresponse. In this study, we found that the expression of miR-34c was significantly decreased in specimens from OS patients with a poor chemoresponse or metastasis compared to those with a good chemoresponse and no metastasis. The inhibition of miR-34c significantly stimulated OS cell invasion and chemoresistance in vitro. In contrast, restoring miR-34c significantly inhibited OS cell invasion and chemoresistance. Furthermore, we identified Notch1 and lymphoid enhancer-binding factor 1 (LEF1) as target genes of miR-34c in OS cells and demonstrated that Notch1 and LEF1 have a major role in the effects of miR-34c on OS cell chemosensitivity and metastasis. Taken together, our data indicate that miR-34c suppresses OS metastasis and chemoresistance by targeting Notch1 and LEF1. Restoring miR-34c may have important implications for the development of strategies for inhibiting metastasis and overcoming OS cell resistance to chemotherapy.

Effects of endostar combined multidrug chemotherapy in osteosarcoma.

Angiogenesis is closely related to tumor development and metastasis. Osteosarcoma is an angiogenesis-dependent tumor, and studies have shown that chemotherapy often induces angiogenesis. Endostatin is a broad spectrum angiogenesis inhibitor and, while pre-clinical trials have shown that the combination of endostatin with chemotherapy can enhance anti-tumor effects, this effect has not yet been shown in clinical trials. Here, we aimed to evaluate the clinical efficacy of endostar (ES, human recombinant endostatin) combined with chemotherapy in the treatment of osteosarcoma patients. A total of 116 newly diagnosed patients with osteosarcoma were enrolled in this study. All patients received 4cycles of chemotherapy with (54 cases) or without (62 cases) ES. ES was administered intravenously at a dose of 15mg/day for 2weeks during each cycle of chemotherapy. The tumors were removed by surgery after 2cycles of chemotherapy treatment, and their histologic response to chemotherapy was evaluated. Immunohistochemistry was used to measure VEGF and CD 31 expression. Chemotherapy increased VEGF expression and the presence of microvessels in osteosarcoma tissues compared with pre-chemotherapy. No significant difference was observed in the histologic response between the ES treatment and non-treatment groups. However, ES treatment significantly inhibited the chemotherapy-induced VEGF expression and presence of microvessels. The ES treatment did not affect the overall survival rate but did increase the event-free survival rate and decreased the occurrence of metastases. In conclusion, our results indicate that antiangiogenic therapy using ES has the potential to prevent the progression of metastases.

IL-17A stimulates the progression of giant cell tumors of bone.

PURPOSE: Giant cell tumors of bone (GCTB) exhibit aggressive bone lytic behavior. Studies have shown that interleukin 17A (IL-17A) is involved pathologic bone resorption in various skeletal disorders. Thus, we have investigated the role of IL-17A in GCTBs. EXPERIMENTAL DESIGN: We evaluated the progression of GCTBs using Campanacci grading and Enneking staging systems in 74 patients with GCTB. The expression of IL-17A and the IL-17A receptor A (IL-17RA) was assessed in GCTB tissues and in both multinucleated giant cells (MNGC) and stromal cells cultured in vitro using immunostaining and reverse transcription PCR (RT-PCR). The effects of IL-17A on the osteolytic activity of the MNGCs and the proliferation of the stromal cells were investigated using the "pit" formation and MTT assays, respectively. The effects of IL-17A on the expression of proosteolytic factors were examined in primary cultured MNGCs and stromal cells using RT-PCR, Western blotting, and gene expression microarrays. RESULTS: In GCTBs, we detected abundant levels of IL-17A, which were associated with tumor extension and grade. IL-17A is predominantly produced by MNGCs, whereas IL-17RA is expressed by both MNGCs and stromal cells in GCTBs. In the MNGCs, the IL-17A increased the mRNA expression of IL-17A and proosteolytic enzymes, and also enhanced osteolytic ability. In the stromal cells, the IL-17A stimulated cellular proliferation and the expression of proosteolytic factors, including RANKL through myc and STAT3, respectively. In addition, IL-17A stimulated in vivo tumor growth and the extent of angiogenesis in GCTBs. CONCLUSION: IL-17A stimulates the progression of GCTBs and might represent a useful candidate marker for progression and as a therapeutic target for GCTBs.

Reconstrução pélvica com aloenxerto ósseo após excisão de tumor / Pelvic reconstruction with allogeneic bone graft after tumor resection

Objetivos: A reconstrução pélvica após excisão de tumor é um desafio. Métodos: realizou-se um estudo retrospectivo para comparar os desfechos entre pacientes submetidos a cirurgia de reconstrução da pelve com aloenxerto ósseo após excisão em bloco de tumores pélvicos e pacientes submetidos apenas à excisão. Resultados: os pacientes sem reconstrução tiveram escores funcionais significantemente menores 3 meses (10 vs. 15, P = 0,001) e 6 meses após a cirurgia (18,5 vs. 22, P = 0,0024), menor tempo de hospitalização (16 dias vs. 40 dias, P < 0,001) e menor custo hospitalar (97.500 vs.193.000 yuans, P < 0,001) do que os que foram submetidos a reconstrução pélvica. Os escores funcionais foram similares 12 meses depois da cirurgia (21,5 vs. 23, P = 0,365) sem diferença na taxa de complicações entre os dois grupos (P > 0,05). Conclusões: A reconstrução pélvica com aloenxerto ósseo depois de cirurgia de tumores pélvicos é associada a desfechos cirúrgicos e funcionais satisfatórios. Outros estudos clínicos são necessários para explorar como selecionar o melhor método de reconstrução. Nível de Evidência IV, Séries de Casos.

Objectives: Pelvic reconstruction after tumor resection is challenging. Methods: a retrospective study had been performed to compare the outcomes between patients undergoing reconstructive surgery of the pelvis with allogeneic bone graft after en bloc resection of pelvic tumors and patients undergoing en bloc resection only. Results: Patients without reconstruction had significantly lower functional scores at 3 months (10 vs. 15, P = 0.001) and 6 months after surgery (18.5 vs. 22, P = 0.0024), a shorter duration of hospitalization (16 days vs. 40 days, P < 0.001), and lower hospital costs (97,500 vs. 193,000 RMB, P < 0.001) than those undergoing pelvic reconstruction. Functional scores were similar after 12 months of surgery (21.5 vs. 23, P = 0.365) with no difference in the rate of complications between the two groups (P > 0.05). Conclusions: pelvic reconstruction with allogeneic bone graft after surgical management of pelvic tumors is associated with satisfactory surgical and functional outcomes. Further clinical studies are required to explore how to select the best reconstruction method. Level of Evidence IV, Case Series.

Pelvic reconstruction with allogeneic bone graft after tumor resection.

OBJECTIVES: : Pelvic reconstruction after tumor resection is challenging. METHODS: A retrospective study had been preformed to compare the outcomes among patients who received pelvic reconstructive surgery with allogeneic bone graft after en bloc resection of pelvic tumors and patients who received en bloc resection only. RESULTS: Patients without reconstruction had significantly lower functional scores at 3 months (10 vs. 15, P = 0.001) and 6 months after surgery (18.5 vs. 22, P = 0.0024), a shorter duration of hospitalization (16 day vs. 40 days, P < 0.001), and lower hospitalization costs (97,500 vs. 193,000 RMB, P < 0.001) than those who received pelvic reconstruction. Functional scores were similar at 12 months after surgery (21.5 vs. 23, P = 0.365) with no difference in the rate of complications between the two groups (P > 0.05). CONCLUSIONS: : Pelvic reconstruction with allogeneic bone graft after surgical management of pelvic tumors is associated with satisfactory surgical and functional outcomes. Further clinical studies are required to explore how to select the best reconstruction method. Level of Evidence IV, Case Series.

Study on the health-related quality of life in patients after surgery for malignant bone tumors.

AIM: We conducted a study in China to assess the health-related quality of life (HRQoL) in patients treated on for malignant bone tumors after surgery, and investigate the possible determinants. METHODS: The subjects were 120 patients surgically treated by amputation and limb-salvage for bone tumors during the period of June 2008 to June 2010. The Medical Outcomes Study Short Form 36 (SF-36) was employed to measure the HRQoL of all the patients before and after surgery. RESULTS: With regard to the results of the general quality of life tool (SF-36), we observed a significant improvement of all the indexes of HRQoL after 6 months (p<0.05). PF, RP and BP scores showed significant increase between surgery after 6 and 12 months (p<0.05). The means of the HRQoL of bone tumor patients in our study were still much lower than those of general population in every domain, even 12 months after surgery. Logistic regression showed that female patients were found to have lower scores in physical component summary (PCS) than males (OR=0.64, 95% CI=0.35-0.89). Patients older than 15 years had lower scores in mental component summary (MCS) (OR=0.60, 95% CI=0.32-0.86). Ablative surgery was related to both lower MCS and PCS scores (For MCS, OR=0.54, 95% CI=0.31-0.83; for PCS, OR=0.43, 95% CI=0.25-0.73). CONCLUSION: Our study showed the treatment for bone tumor could greatly alter the HRQoL of patients. Age, sex and type of surgery were associated with physical or mental HRQoL after surgery.

Magnetic resonance imaging is appropriate for determining the osteotomy plane for appendicular osteosarcoma after neoadjuvant chemotherapy.

There are no standard criteria for determining a sufficient resection margin in the treatment of osteosarcoma. The purposes of this study are to evaluate clinical outcomes using T1-weighted magnetic resonance imaging (MRI) for determining the margin of resection and to compare that with the results of different imaging modalities. Seventeen patients diagnosed with osteosarcoma who underwent en bloc resection with a margin of 2-3 cm based on T1-weighted MRI following chemotherapy were studied. Imaging modalities including conventional radiography, MRI, computed tomography (CT), visual assessment, and histopathological examination were performed and compared. Survival rates were determined. After follow-up of 45.5 ± 13.8 months, no local tumor recurrence was observed in any patient. The 1-, 3-, and 5-year survival rates were 94.1, 82.3, and 76.5%, respectively. The differences in the measurement errors among the five methods were analyzed using pathology as the gold standard. Errors were smallest using T1-weighted and fat-suppressed MRI. There were no significant differences between the measurement results of postoperative histopathological examination and that of T1-weighted imaging or T2 fat-suppressed imaging. The measurement results of radiography and CT were significantly different from that of postoperative pathological findings (P < 0.05). Thus, MRI examination is superior to radiography and CT for determining tumor invasion in patients with osteosarcoma. A resection margin of 2-3 cm determined by MRI provides adequate treatment, while minimizing tissue removal.