Enneking Stage Research Articles

EFEMP1 as a Potential Biomarker for Diagnosis and Prognosis of Osteosarcoma.

Osteosarcoma (OS) is the most common primary bone malignancy. Our previous study revealed an association between the level of epidermal growth factor-containing fibulin-like extracellular matrix protein 1 (EFEMP1) and the invasion, metastasis, and poor prognosis of OS. However, the exact correlation between the serum EFEMP1 level and OS diagnosis and progression was unclear. This study is aimed at determining the value of the serum EFEMP1 level in the diagnosis and prognosis of OS. Fifty-one consecutive OS patients were prospectively registered in this study. The serum EFEMP1 levels were measured using ELISA at diagnosis, after neoadjuvant chemotherapy, and before and after surgical treatment. Sixty-nine healthy subjects in the control group, nine patients with chondrosarcoma, and 12 patients with giant cell tumor of the bone were also enrolled in this study. Surgical orthotopic implantation was used to generate a mouse OS model, and the correlation between the circulating EFEMP1 levels and tumor progression was examined. Then, OS patients had significantly higher mean serum EFEMP1 levels (7.61 ng/ml) than the control subjects (1.47 ng/ml). The serum EFEMP1 levels were correlated with the Enneking staging system (r = 0.32, P = 0.021) and lung metastasis (r = 0.50, P < 0.001). There was also a correlation between the serum EFEMP1 level and EFEMP1 expression in the respective OS samples (r = 0.49, P < 0.001). Additionally, patients with either chondrosarcoma or giant cell tumor of the bone had significantly higher serum EFEMP1 levels than OS patients. Surgical and chemotherapeutic treatment led to an increase in the serum EFEMP1 levels. Then, the destruction of bone tissues might be one of the factors about the EFEMP1 levels. In the mouse OS model, the serum EFEMP1 level was correlated with tumor progression. Our results suggested that serum EFEMP1 levels might be used to distinguish OS patients from healthy controls and as an indicator for OS lung metastasis. Serum EFEMP1 levels could serve as a new and assisted biomarker for the auxiliary diagnosis and prognosis of OS.

A four-factor immune risk score signature predicts the clinical outcome of patients with spinal chordoma.

BackgroundCurrently, the measurement of immune cells in previous studies is usually subjective, and no immune‐based prognostic model has been established for chordoma. In this study, we sought to simultaneously measure tumor‐infiltrating lymphocyte (TIL) subtypes in chordoma samples using an objective method and develop an immune risk score (IRS) model for survival prediction.MethodsMultiplexed quantitative immunofluorescence staining was used to determine the TIL levels in the tumoral and stromal subareas of 114 spinal chordoma specimens (54 in the training and 60 in the validation cohort) for programmed death‐1 (PD‐1), CD3, CD8, CD20 (where CD is cluster of differentiation), and FOXP3. Flow cytometry was performed to validate the immunofluorescence assay for lymphocyte measurement on an additional five fresh chordoma specimens. Subsequently, the IRS model was built using the least absolute shrinkage and selection operator (LASSO) Cox regression method.ResultsFlow cytometry and quantitative immunofluorescence showed similar lymphocytic percentages and TIL subpopulation proportions in the fresh tumor specimens. With the training data, the LASSO model identified four immune features for IRS construction: tumoralFOXP3, tumoralPD‐1, stromalFOXP3, and stromalCD8. In both cohorts, a high IRS was significantly associated with tumoral programmed cell death‐1 ligand 1 expression, Enneking inappropriate tumor resection, and surrounding muscle invasion by tumor. Multivariate Cox regression and stratified analysis in the two cohorts revealed that the IRS was an independent predictor and could effectively separate patients with similar Enneking staging into different risk subgroups, with significantly different survival rates. Further receiver operating characteristic analysis found that the IRS classifier had a better prognostic value than the traditional clinicopathological factors and compensated for the deficiency of Enneking staging for outcome prediction. More importantly, a nomogram based on the IRS and clinical predictors showed adequate performance in estimating disease recurrence and survival of patients.ConclusionsThese data support the use of the IRS signature as a reliable prognostic tool in spinal chordoma and may facilitate individualized therapy decision making for patients.

Prognostic value of pre-treatment Naples prognostic score (NPS) in patients with osteosarcoma

BackgroundThis study aimed to evaluate the clinical significance of pre-treatment Naples prognostic score (NPS) in patients with osteosarcoma.MethodsThe clinical data of 133 osteosarcoma patients between January 2011 and February 2018 in our hospital was retrospectively collected and analyzed. NPS was calculated from four parameters, including serum albumin level, serum total cholesterol (TC), lymphocyte-to-monocyte ratio (LMR), and neutrophil-to-lymphocyte ratio (NLR). Patients were divided into three groups (group 1-3) based on NPS. The relationships between NPS and clinical features, overall survival (OS), and progression-free survival (PFS) were analyzed. Two prediction models based on NPS and clinical parameters were developed: clinical parameters model (model A), and the combined model of NPS and clinical parameters (model B). Their predictive performances were further evaluated and compared.ResultsThe median follow-up time of this cohort was 46.0 (range, 5–75) months, while the median OS and PFS was 40 (range, 5–75) months and 36 (range, 5–71) months, respectively. NPS was significantly correlated with gender, tumor location, Enneking stage, pathological fracture, local recurrence, and metastasis (all P < 0.05). Variables of NPS, Enneking stage, local recurrence, metastasis, and NLR were confirmed as independent prognostic factors for OS and PFS by univariate and multivariate Cox analysis. Prediction model B obtained larger AUCs for OS and PFS and showed better consistency between nomogram-predicted and actual survival than that of model A at the follow-up time of 1-, 3-, and 5-year.ConclusionsNPS was a novel, reliable, and multidimensional prognostic scoring system with favorable predictive performance for patients with osteosarcoma.

Silencing of RIPK4 inhibits epithelial‑mesenchymal transition by inactivating the Wnt/β‑catenin signaling pathway in osteosarcoma.

Receptor interacting protein kinase 4 (RIPK4) is a serine/threonine kinase that plays an important role in the regulation of cell proliferation, invasion and metastasis in several malignancies; however, its clinical significance and biological function in osteosarcoma (OS) remains unknown. In the present study, the RIPK4 expression level was significantly upregulated in OS tissues and cell lines. High RIPK4 expression was positively associated with larger sized tumors, advanced Enneking stage and poor prognosis in patients with OS. Furthermore, the results revealed that RIPK4 knockdown in the OS cell lines MG-63 and U2OS reduced cell migration and invasion via the inhibition of epithelial-mesenchymal transition (EMT) process, whereby E-cadherin expression was increased and N-cadherin and vimentin expression decreased. Mechanistically, RIPK4 knockdown inhibited EMT by inactivating the Wnt/β-catenin signaling pathway. These findings suggest that RIPK4 may be a novel potential therapeutic target for the treatment of metastases in patients with OS.

Expression Changes, Prognostic Analysis and Risk Factors of miR-625-3p and miR-449a in Osteosarcoma Patients after Surgery

The expression changes of miR-625-3p and miR-449a in osteosarcoma patients after surgery is the critical study in this paper. Analysis of their prognosis and risk factors can be helpful in understanding the prognostic value for clinical purposes. Fifty-eight patients with osteosarcoma diagnosed in our hospital were considered as the research group (RG), and 52 health subjects at the same time were collected as the control group (CG). Fluorescence quantitative PCR (RTPCR) was employed to test the expression levels of miR-625-3p and miR-449a in the serum of subjects in both groups before surgery and patients in the RG after surgery. The survival conditions of both groups with high and low miR-625-3p and miR-449a expression levels were compared after 5-year follow-up of patients in the RG. The prognostic factors of osteosarcoma patients were assessed through Cox regression analysis. The miR-625-3p expression level in the RG was dramatically higher than that in the CG (p < 0.05), and the miR-449a expression level was dramatically lower than that in the CG (p < 0.05). The miR-625-3p expression level in the RG after surgery was markedly lower than that before surgery (p < 0.05), and the miR-449a expression level was markedly higher than that before surgery (p < 0.05). After they were followed up for 5 years, the overall survival rate of those in the RG was 60.34%, the rate of miR-625-3p low expression group (LEG) was dramatically higher than that of high expression group (HEG) (p < 0.05), and the rate of miR-449a HEG was markedly higher than that of miR-449a LEG (p < 0.05). The relative expression of miR-625-3p and miR- 449a was remarkably different from tumor size, lymph node metastasis, and Enneking stage (p < 0.05). Univariate Cox regression analysis identified that tumor size, lymph node metastasis, Enneking stage, miR-625-3p and miR-449a were risk factors affecting the prognosis of patients; multivariate Cox regression analysis found that the five were independent prognostic factors of them. miR-625-3p and miR-449a are differentially expressed in the postoperative serum of osteosarcoma patients, and their differential expression is an independent risk factor affecting their prognosis, which has prognostic value and can provide a reference value for clinical practice.

Factors Affecting Prognosis of Patients With Osteoblastoma of the Mobile Spine: A Long-Term Follow-up Study of 70 Patients in a Single Center.

Osteoblastoma (OBL) in the mobile spine is a benign tumor with aggressive potential. There is not much published information on OBL of the mobile spine as a result of rarity of the disease. There are controversies over the aggressive subtype and prognostic factors of the condition. To explore the aggressive subtype and investigate prognostic factors of OBLs in the mobile spine. A retrospective analysis of spinal OBL was performed by survival analysis. Patients in 2 cohorts were analyzed based on the Enneking grade and pathological evaluation. Local recurrence-free survival (RFS) was estimated using the Kaplan-Meier method to identify potential prognostic factors. Factors with P ≤.05 were subjected to multivariate analysis by Cox regression analysis. P ≤.05 were considered statistically significant. Included in this study were 70 patients with spinal OBL who were followed up by a mean of 67.1 mo (range 24-188). Local recurrence was detected in 9 cases after initial surgery, and death occurred in 2 cases. The result of statistical analysis suggested that Epithelioid OBL (EO) with Enneking stage 3 (St.3) and total spondylectomy were independent prognostic factors for RFS. St.3 or EO lesions seem to be more aggressive than St.2 or conventional osteoblastomas, but St.3 and EO should be considered simultaneously in predicting the aggressiveness of the lesion and the risk of recurrence. Total spondylectomy performed either by en bloc or piecemeal could significantly reduce recurrence of OBLs in the mobile spine.

Correlation between the changes of serum COX 2, APE1, VEGF, TGF-β and TSGF levels and prognosis in patients with osteosarcoma before and after treatment.

Osteosarcoma (OS) is a progressive primary bone tumor that originates from immature stromal spindle cells. After chemotherapy, the serum-related indexes which are related to the prognosis. The aim of this study is to investigate the correlation between changes in serum cyclooxygenase-2 (COX-2), basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), transforming growth factor-beta (TGF-β), and tumor-specific growth factor (TSGF) levels and prognosis of patients with osteosarcoma (OS) before and after treatment. Data of 75 patients with OS (observation group) and 55 healthy controls (control group) were retrospectively analyzed. Chemotherapy was administered to the observation group. Serum lactate dehydrogenase, alkaline phosphatase, and TSGF levels were measured before and after treatment. The observation group patients were classified as normal or abnormal according to the changes in serum COX-2, bFGF, VEGF, TGF-β, and TSGF levels after chemotherapy. Patients were followed up for 7.5 years, and the survival rate was determined. Single-factor influencing prognosis was included in the Cox model, and independent factors influencing prognosis were analyzed. After chemotherapy, the mean serum COX-2, bFGF, VEGF, and TSGF levels decreased significantly in the observation group but were still higher than those in the control group. Furthermore, serum TGF-β levels increased in the observation group but were still lower than those in the control group. The 5-year survival rate of patients with normal serum COX-2, bFGF, VEGF, and TSGF levels was significantly higher in the normal subgroup than in the abnormal subgroup. Cox analysis showed that the Enneking stage and COX-2 level after chemotherapy were independent prognostic factors. The serum COX-2, bFGF, VEGF, and TSGF levels of patients with OS significantly changed after chemotherapy, and the short-term survival rate of patients with normal levels of these biomarkers after chemotherapy was high.

Upregulated lncRNA THRIL/TNF-α Signals Promote Cell Growth and Predict Poor Clinical Outcomes of Osteosarcoma.

BackgroundThe immunosuppressive facet and tumorigenic role of TNF-α have been revealed in osteosarcoma (OS). Long noncoding RNA THRIL is identified to regulate TNF-α expression and participates in immune response. Thus, investigations on the clinical expression pattern of THRIL/TNF-α signal in OS would provide a potential target premise for OS patients.MethodsWe collected OS (n=83), nontumor tissues (n=37) and serum samples (n=83 for OS and n=40 for healthy control) to determine the expressions and clinical significance of THRIL/TNF-α signal. Knockdown of THRIL in OS cell lines MG63 and Saos2 in vitro and in vivo was performed to confirm its function in the development of OS.ResultsElevated expression of THRIL was associated with increased TNF-α levels in OS tissues and serum samples. Combination of THRIL and TNF-α in tissues showed a more efficient diagnostic value for OS patients than either of them. Moreover, high-expressed THRIL was associated with larger tumor size, advanced Enneking stage and lung metastasis, whereas high TNF-α expression was found in patients with high histologic grade and patients who simultaneously harbor high THRIL and TNF-α showed the worst overall survival and metastasis-free survival. TNF-α signals increased OS cell vitalities in vitro but knockdown of THRIL inhibited TNF-α expressions, leading to impaired cell vitality, increased apoptosis and also downregulated epithelial to mesenchymal transition (EMT) phenotype and the ability of invasion, but these processes were restored by the treatment of TNF-α. The oncogenic role of THRIL/TNF-α signal was also confirmed in the xenograft model of MG63 cells.ConclusionOverexpressed THRIL and TNF-α promoted OS progression with efficient diagnostic and prognostic value. THRIL/TNF-α signal supported cell growth and EMT phenotype of OS cells in vitro and in vivo.

High level of RNF187 contributes to the progression and drug resistance of osteosarcoma.

Objectives: Ring finger protein 187 (RNF187) was recently demonstrated to be up-regulation and function as a promoter in multiple cancers. However, the roles of RNF187 in osteosarcoma (OS) are unclear. Here, we tried to reveal the clinicopathological and biological roles of RNF187 in OS.Materials and Methods: We employed the quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC) to determine the expression of RNF187 in OS tissues and cells. Migration and invasion capacities were analyzed by wound healing and transwell assays, and colony formation and CCK8 assays were performed to investigate proliferative ability. The functional effects of RNF187 on OS drugs resistance were further determined by CCK8 and western blot assays. Then, the relationship between RNF187 expression and clinical implications was analyzed by tissue microarrays (TMAs) including 51 OS cases. Moreover, the prognostic value was also determined by Kaplan-Meier analysis.Results: We reported that RNF187 mRNA was significantly increased in OS tissues compared to matched nontumorous tissues (3.83 ±0.79 vs. 1.70 ± 0.63), which was in line with the IHC assay in TMAs. By RNA interference and cDNA transfection, we showed high level of RNF187 increased the migration, invasion and proliferation of OS cells. Moreover, we demonstrated that elevated RNF187 expression induced OS cell drugs resistance, activated the ERK1/2 molecular and markedly enhanced the BCL-2 expression. Clinically, OS patients with high level of RNF187 was associated with Histologic differentiation (p=0.001), an advanced Enneking stage (p=0.001), response to chemotherapy (p=0.004), and metastasis (p= 0.001). Clinically, our data displayed that the RNF187 overexpression in OS samples associated with shorten overall survival (p=0.001) and high tumor recurrence (p=0.001) in postoperative OS patients.Conclusions: Our results indicate that Elevated RNF187 expression is a new adverse outcomes marker for OS patients and may be used as a new therapeutic target of OS.

LncRNA KRAL suppresses cell growth and increases sensitivity to doxorubicin in osteosarcoma cells by sponging microRNAs-141

Osteosarcoma (OS) is one of the most common types of malignant tumors characterized by uncontrolled proliferation ability and acquired drug resistance. The previous study indicated that lncRNA KRAL participated in the reversal of 5-FU resistance in liver cancer, but it remains unclear whether lncRNA KRAL involved in doxorubicin (DOX) resistance of osteosarcoma. The expression of lncRNA KRAL and MicroRNAs-141 (miR-141) were detected by RT-qPCR experiment. Also, we used the plasmids transfection to construct the lncRNA KRAL overexpressed OS cell lines. Subsequently, the cell proliferation ability and the sensitivity to DOX in OS cells upon upregulating lncRNA KRAL expression were analyzed via CCK-8 and EDU assay, while western blotting experiment was performed to detect the regulatory mechanism. We found that lncRNA KRAL was downregulated in OS tissues, and the OS patients with OS patients with lower expression of lncRNA KRAL were more likely to have advanced Enneking stage, larger tumor size and distant metastasis. Subsequently, we discovered that upregulation of lncRNA KRAL could inhibit cell proliferation and increase the sensitivity to DOX of OS cells. Interestingly, the western blot results showed that over-expression of lncRNA KRAL could lead to down-expression of P-gp protein and reversal of Epithelial–mesenchymal transition (EMT) pathway. Furthermore, we identified miR-141 as the downstream target gene of lncRNA KRAL, which was further confirmed by the luciferase reporter assay. More importantly, our data demonstrated that addition of miR-141 could reverse cell proliferation and drug sensitivity of lncRNA KRAL-overexpressed OS cells. LncRNA KRAL could suppress cell growth and increases sensitivity to DOX in OS cells by sponging miR-141.

Hsa_circ0021347 as a Potential Target Regulated by B7-H3 in Modulating the Malignant Characteristics of Osteosarcoma.

In our previous study, we showed that B7-H3 played crucial roles in osteosarcoma (OS) development and might serve as a negative regulator of in osteoimmunology and help tumor cells escape immune surveillance. However, little is known about B7-H3 deficiency and its corresponding circRNA alteration or their relationship with osteosarcoma progression. Therefore, we established stable silencing of B7-H3 in OS cells and validated our results with western blotting and real-time PCR detection. Then, we performed a circRNA array to analyze the differential expression of circRNAs between the control and B7-H3 knockdown cells. The association between target circRNA expression and the clinicopathological features of patients with OS was further analyzed. As a result, hsa_circ0021347 was selected and validated to be significantly downregulated in OS tissues and cell lines and showed a strong negative relationship with B7-H3 expression in OS. In addition, clinicopathological features showed that hsa_circ0021347 in OS tissues was negatively associated with Enneking stage and positively associated with patients' survival. Finally, Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and PANTHER pathway analyses were performed to predict a network of hsa_circ0021347/miRNAs interactions to help us develop potential biomarkers for clinical diagnosis and design therapeutic strategies for OS.

Application of pasteurized tumor-bearing bone replantation for primary malignant bone tumor of extremities

To assess the effectiveness of the pasteurized tumor-bearing bone replantation in treatment of primary malignant bone tumor of extremities. Between February 2012 and June 2016, 13 patients with primary malignant bone tumor of extremities were treated with pasteurized tumor-bearing bone replantation after extensive excision. There were 8 males and 5 females, aged from 11 to 27 years, with an average of 17.4 years. Tumors were located at the mid-upper humerus in 2 cases, the mid-upper femur in 2 cases, the mid-lower femur in 6 cases, the mid-upper tibia in 2 cases, and the middle tibia in 1 case. According to Enneking staging system, 3 patients were classified as stage ⅠB, 5 patients as stage ⅡA, and 5 patients as stage ⅡB. There were 11 cases of osteosarcoma and 2 cases of chondrosarcoma. The disease duration ranged from 3 to 8 months, with an average of 4.8 months. The length of the tumor-bearing bone ranged from 8 to 16 cm, with an average of 12.5 cm. Postoperative follow-up was conducted regularly to evaluate the status of inactivated bone and complications. The limb function was assessed by the Musculoskeletal Tumor Society 93 (MSTS-93) scoring system. All 13 patients were followed up 26-79 months, with an average of 50.5 months. Eleven patients survived without tumors. Two patients had local soft tissue recurrence and underwent amputation, 1 had lung metastasis after amputation and died. All patients had no complications of bone resorption, shortening, and internal fixation disorder. The nonunion of osteotomy occurred in 4 cases. Among them, 3 cases were successfully healed after autologous iliac bone grafting, and 1 case was treated conservatively. The healing time of metaphyseal osteotomy end was 10-15 months (mean, 12.6 months), the healing rate was 90.9% (10/11); the healing time of diaphyseal osteotomy end was 12-21 months (mean, 17.0 months), the healing rate was 72.7% (8/11); and the total healing rate of osteotomy end was 81.8% (18/22). One case had inactivated bone fracture and 1 case had incision dehiscence and infection after operation. At last follow-up, the MSTS-93 score of affected limb ranged from 21 to 28, with an average of 25.3. The procedure of the pasteurized tumor-bearing bone replantation is an effective, simple, and economic way in repair of massive segmental bone defect to save limb function for primary malignant bone tumor of extremities.

Association of Mental Health-Related Proteins DAXX, DRD3, and DISC1 With the Progression and Prognosis of Chondrosarcoma.

Chondrosarcoma is the second most common malignant bone tumor. Current therapies remain ineffective, resulting in poor prognoses. Biomarkers for chondrosarcoma and predictors of its prognosis have not been established. Mental health–related proteins have been associated with the pathogenesis, progression, and prognosis of many cancers, but their association with chondrosarcoma has not been reported. In this study, the expression and clinicopathological significance of the mental health–related proteins DAXX, DRD3, and DISC1 in chondrosarcoma tissue samples were examined, over an 84-months follow-up period. In immunohistochemical analysis, the rates of positive DAXX, DRD3, and DISC1 expression were significantly higher in chondrosarcoma than in osteochondroma tissue (P < 0.01). The percentages of positive DAXX, DRD3, and DISC1 expression were significantly lower in tissues with good differentiation (P < 0.01), AJCC stage I/ II (P < 0.01), Enneking stage I (P < 0.01), and non-metastasis (P < 0.05), respectively. In Kaplan–Meier survival analysis, significantly shorter mean survival times were associated with moderate and poor differentiation (P = 0.000), AJCC stage III/IV (P = 0.000), Enneking stage II/III (P = 0.000), metastasis (P = 0.019), invasion (P = 0.013), and positive DAXX (P = 0.012), and/or DRD3 (P = 0.018) expression. In Cox regression analysis, moderate and poor differentiation (P = 0.006), AJCC stage III/IV (P = 0.013), Enneking stage II/III (P = 0.016), metastasis (P = 0.033), invasion (P = 0.011), and positive DAXX (P = 0.033), and/or DRD3 (P = 0.025) staining correlated negatively with the postoperative survival rate and positively with mortality. In competing-risks regression analysis, differentiation (P = 0.005), metastasis (P = 0.014), invasion (P = 0.028), AJCC stage (P = 0.003), Enneking stage (P = 0.036), and DAXX (P = 0.039), and DRD3(P = 0.019) expression were independent predictors of death from chondrosarcoma. The areas under receiver operating characteristic curves for DAXX, DRD3, and DISC1 expression were 0.673 (95% CI, 0.557–0.788; P = 0.010), 0.670 (95% CI, 0.556–0.784; P = 0.011), and 0.688 (95% CI, 0.573–0.802; P = 0.005), respectively. These results suggest that DAXX, DRD3, and DISC1 could serve as biomarkers of chondrosarcoma progression and predictors of its prognosis.

Association of high PDPN expression with pulmonary metastasis of osteosarcoma and patient prognosis.

Podoplanin (PDPN) is an important positive regulator of platelet aggregation and functions as a lymphatic endothelial marker. PDPN has been observed to be expressed in human tumor tissues and various cancer cell lines. In the present study, PDPN expression in patients with primary osteosarcoma was assessed at the mRNA and protein levels, and the associations between PDPN expression and pulmonary metastasis (PM) and prognosis were examined. Reverse transcription-quantitative PCR (RT-qPCR) analysis was used to detect the expression levels of PDPN in primary osteosarcoma tissues and paired normal bone tissues (n=20 pairs). In addition, immunohistochemical analysis of PDPN expression was performed in 168 paraffin-embedded osteosarcoma tissue specimens and 23 matched normal tissues. The RT-qPCR results revealed higher mRNA expression levels of PDPN in patients with PM compared with patients without PM. Further survival analyses identified Enneking stage and PM as two independent prognostic indicators. Finally, univariate analysis revealed that high PDPN protein expression was significantly associated with Enneking stage and PM in patients with osteosarcoma.

Treatment of Typical Enneking Stage 3 Thoracic Aggressive Vertebral Hemangiomas with Pain and Neurologic Deficits: Results After at Least 36 Months of Follow-Up

To evaluate the efficiency of a comprehensive treatment strategy for patients with Enneking stage 3 thoracic aggressive vertebral hemangioma (AVH). A retrospective analysis of radiographic and clinical outcomes was performed for 17 patients who received treatment for Enneking stage 3 thoracic AVH accompanied by pain and neurologic deficits between January 2010 and February 2015. A visual analog scale (VAS) was used to clinically evaluate the level of pain. Neurologic examinations were performed to assess the patients' sensory symptoms, motor deficits, and Frankel grade. The average operative time was 181.8 ± 37.1 minutes, and the average intraoperative blood loss was 1226.5 ± 151.2 mL (range, 900-1450 mL). All patients underwent preoperative embolization to minimize intraoperative blood loss. All patients experienced immediate pain relief and resolution of their neurologic symptoms. All 17 patients achieved Frankel grade D at the final follow-up; moreover, they reported that their pain was relieved (mean VAS score, 2.82 ± 0.81; P < 0.05) and their neurologic deficits had resolved. No surgery-related complications were noted. No patients exhibited signs of recurrence. We recommend a comprehensive treatment strategy for patients with Enneking stage 3 thoracic AVH that includes preoperative embolization, vertebroplasty, posterior decompression, and internal fixation. We recommend that absorbable gelatin sponge particles be used for intraoperative embolization prior to the injection of bone cement, which may significantly reduce intraoperative bleeding, operative time, and occurrence of surgery-related complications.

Circular RNA LARP4 correlates with decreased Enneking stage, better histological response, and prolonged survival profiles, and it elevates chemosensitivity to cisplatin and doxorubicin via sponging microRNA-424 in osteosarcoma.

BackgroundThis study aimed to evaluate the association of circular RNA La‐related RNA‐binding protein 4 (circ‐LARP4) with clinical features and prognosis in osteosarcoma patients, and further explore its effect on chemosensitivity in osteosarcoma cells.MethodsSeventy‐two osteosarcoma patients with Enneking stage IIA‐IIB who underwent resection were consecutively enrolled, and then, tumor tissues and non‐tumor tissues were obtained. Circ‐LARP4 in tumor tissue/non‐tumor tissue was detected by quantitative polymerase chain reaction. After circ‐LARP4 overexpression and negative control overexpression plasmid transfection, relative cell viability (%) was evaluated by Cell Counting Kit‐8 in MG63 cells treated by different concentrations of cisplatin, methotrexate, and doxorubicin, and IC50 was calculated.ResultsCirc‐LARP4 was downregulated in tumor tissue compared with non‐tumor tissue and had a good value in distinguishing tumor tissue from non‐tumor tissue with an area under curve of 0.829 (95% CI: 0.762‐0.859). Meanwhile, tumor circ‐LARP4 was negatively correlated with the Enneking stage. After resection, circ‐LARP4 high expression patients showed an increased tumor cell necrosis rate to adjuvant chemotherapy compared to circ‐LARP4 low expression patients, and circ‐LARP4 high expression correlated with prolonged disease‐free survival and overall survival. In vitro experiments revealed that circ‐LARP4 overexpression elevated the chemosensitivity of MG63 cells to cisplatin and doxorubicin but not methotrexate, with decreased cisplatin IC50 and doxorubicin IC50 concentrations than negative control. Besides, miR‐424 overexpression attenuated the chemosensitivity in circ‐LARP4 overexpression‐treated MG63 cells.ConclusionCirc‐LARP4 high expression correlates with decreased Enneking stage and prolonged survival profiles, and it elevates chemosensitivity to cisplatin and doxorubicin via sponging miR‐424 in osteosarcoma.

Dysregulation of microRNA-106a-5p-RUNX1 axis associates with clinical progression and prognosis of osteosarcoma patients

MicroRNA-106a-5p (miR-106a-5p) functions as a tumor suppressor in osteosarcoma cells. Here, we aimed to identify novel target genes of miR-106a-5p in osteosarcoma, as well as to investigate their prognostic value and the biological functions. At first, the mammalian runt-related factor 1 (RUNX1) was identified as one of the target genes of miR-106a-5p in osteosarcoma cells by luciferase reporter gene assay, real-time quantitative RT-PCR and Western blot analysis. Then, the expression levels of miR-106a-5p and RUNX1 in osteosarcoma tissues were detected, and their associations with clinicopathological features and patients' prognosis were statistically analyzed. Compared with adjacent non-cancerous tissues, miR-106a-5p and RUNX1 mRNA/protein expression in osteosarcoma tissues were significantly decreased and increased, respectively (all P < 0.01). Low miR-106a-5p, high RUNX1 and miR-106a-5p-low/RUNX1-high expression in osteosarcoma tissues were all significantly associated with advanced Enneking stage, positive metastasis and shorter overall survival (all P < 0.05). Moreover, miR-106a-5p and RUNX1 expression, alone or in combination, were identified as independent prognostic factors for osteosarcoma patients' overall survival. Functionally, the enforced expression of miR-106a-5p significantly suppressed proliferation and invasion of osteosarcoma cells, while the overexpression of RUNX1 effectively reversed its suppressive roles. In conclusion, our findings show the dysregulation of miR-106a-5p-RUNX1 axis in human osteosarcoma tissues and suggest its crucial roles in cancer progression and patients' prognosis. More interestingly, miR-106a-5p may function as a tumor suppressor in osteosarcoma cells via regulating its target gene RUNX1.

Osteoclastoma of bone: Role of extended curettage with allogenic bone grafting

Background: Mesenchymal stromal cells are origin for benign tumor osteoclastoma of bone. Different location of presentations with varied manifestations is a unique feature of osteoclastoma. Extended curettage of the lesion and supplementary allogenic bone grafting in selected cases is widely used treatment regimen. Selection of cases is important for successful outcome using this as a treatment protocol. Our study with 30 cases aims in selection of cases for successful outcome with this treatment protocol. Materials and Methods: Biopsy proven (core and open biopsy) 30 cases of osteoclastoma are selected. The cases are evaluated clinically, radiographically, CT &MRI scan and by histology. Campanancci grading and Enneking staging was used in the study. Chi square tests, Mann Whitney test and ANOVA were used for statistical analysis. Tumors with cortical breach which is confined to one surface and cortical breach less than one third of circumference were managed by extended curettage and bone grafting.Results: The average age of presentation was 25 years. Distal femur is the commonest site (16 cases) followed by upper end of tibia (12 cases) and 2 cases are from distal radius. Pathological fractures were seen in 2 cases. 4 tumors had less than 5 mm of subchondral bone free of tumor. The average follow up period was 18.6 months (range 2-84). 4 cases of recurrence were seen in a grade III recurrent distal femur lesion (3) and proximal tibia group (1) in the intralesional curettage group.Conclusion: Extended curettage with allogenic bone grafting is a reliable method with good functional outcome with less complications than other methods (enbloc excision) whenever we stick on to principles of tumor surgery with proven guidelines of case selection. Recurrence can always be treated with recurettage.

Roles Of EAAT1, DHFR, And Fetuin-A In The Pathogenesis, Progression And Prognosis Of Chondrosarcoma.

AimsChondrosarcoma (CS) is a high-morbidity, relatively common bone malignancy without well-established biomarkers. The proteins EAAT1, DHFR, and fetuin-A have been investigated in many cancers, but their specific relationship to CS has not been reported. The present study examined EAAT1, DHFR, and fetuin-A expression in CS and the clinicopathological significance of these proteins in CS pathogenesis, progression, and prognosis.MethodsEAAT1, DHFR, and fetuin-A protein levels in 80 CS and 25 chondroma specimens were measured by immunohistochemistry and related to histological and clinical factors with chi-squared tests. Following univariate survival analysis, ROC curves calculation, and multivariate analysis.ResultsEAAT1, DHFR, and fetuin-A expression levels were significantly higher in the CS group than in the chondroma group (p < 0.05). Their immunopositivity rates were significantly greater in tissues with moderate or poor tumor differentiation, AJCC stage III or IV, Enneking stage II or III, and metastasis (p<0.05 or p<0.01 or p<0.001). Kaplan–Meier survival analysis showed significantly shorter survival in patients with moderately or poorly differentiated tumors, AJCC stage III or IV CS, Enneking stage II or III CS, metastasis, invasion, or EAAT1, DHFR, and fetuin-A immunopositivity (p < 0.05 or p < 0.001). Cox regression analysis showed that moderate or poor tumor differentiation, AJCC stage III or IV, Enneking stage II or III, metastasis, invasion, and EAAT1, DHFR, or fetuin-A immunopositivity correlated negatively with postoperative survival and positively with mortality (p < 0.05). The AUCs for EAAT1, DHFR, and fetuin-A were 0.654 (95% CI: 0.532–0.776, p = 0.025), 0.638 (95% CI: 0.519–0.756, p = 0.039), and 0.670 (95% CI: 0.556–0.784, p = 0.011), respectively.ConclusionEAAT1, DHFR, and fetuin-A may be important biomarkers of the pathogenesis and progression of CS and predictors of its prognosis.

Transferrin receptor-1 and VEGF are prognostic factors for osteosarcoma

BackgroundOsteosarcoma is aggressive and prognostic biomarkers are important to predict the outcomes of surgery and chemotherapy. Here, we investigated the potential of transferrin receptor-1 (TfR1) and vascular endothelial growth factor (VEGF) as prognostic markers of osteosarcoma.MethodsTfR1 and VEGF in osteosarcoma samples from a cohort of 53 osteosarcoma patients were detected by immunohistochemistry analysis. The correlation of TfR1 and VEGF levels with clinicopathological parameters was analyzed by Pearson chi-square and Spearman-rho tests. Overall patient survival was analyzed by the Kaplan-Meier method.ResultsWe found that TfR1 and VEGF expression levels were low in 20.8% and 18.9%; modest in 35.8% and 35.8%; and high in 43.4% and 45.3% of osteosarcoma patients, respectively. TfR1 and VEGF expression was significantly correlated to histologic grade, Enneking stage, and distant metastasis. TfR1 expression was significantly correlated to VEGF expression and both TfR1 expression and VEGF expression were correlated to shorter overall survival.ConclusionsTfR1 and VEGF are potential prognostic factors for osteosarcoma.