巨噬细胞移动因子在骨肉瘤患者血清中的表达及临床意义

[目的]评价巨噬细胞移动因子(MIF)在骨肉瘤患者血清中的表达及临床意义。[方法]应用酶联免疫吸附分析(ELISA)方法,检测30名健康人(对照组)和32例骨肉瘤患者(病例组)初次入院、术后化疗前、术后化疗结束时血清MIF水平,分析MIF的表达与骨肉瘤临床病理特征的关系以及化疗对MIF水平的影响。[结果]病例组患者入院时血清MIF为(1.521±0.520)ng/ml,显著高于对照组的(1.149±0.284)ng/ml(t=3.452,P=0.001)。MIF的血清水平与骨肉瘤临床分期、肿瘤大小和转移存在相关性。患者化疗前血清中MIF水平显著高于化疗后(P<0.05)。[结论]MIF在骨肉瘤患者血清中高表达,可能为骨肉瘤的重要预后因子及潜在的治疗靶点。     

血小板反应蛋白-1的表达与骨肉瘤新生血管生成的研究

目的：检测血小板反应蛋白-1（thrombospodin-1,TSP-1）在骨肉瘤中的表达,探讨其与骨肉瘤组织血管生成、侵袭性的关系。方法：收集2003年1月-2007年2月手术切除的54例骨肉瘤标本,应用免疫组化法,检测骨肉瘤组织中TSP-1的表达,CD34单克隆抗体标记血管内皮细胞计数骨肉瘤微血管密度（mi—eI＇ovesse]density,MVD）。结果：骨肉瘤TSP-1的表达程度与MVD呈负相关,TSP-1高表达组的MVD明显低于TSP-1低表达组;骨肉瘤TSP-1表达与骨肉瘤临床病理因素无相关性,但与肺转移显著负相关。结论：TSP-1在骨肉瘤发生发展中起重要作用,并影响血管形成及肿瘤侵袭性,有望成为治疗骨肉瘤的新靶点。     

Prognostic Significance of Serum Lactate Dehydrogenase in Patients with Osteosarcoma of the Extremities

Summary

Six hundred and fifty-six patients with osteosarcoma of the extremities (107 metastatic and 549 with localized disease) were followed from 2.5 to 20 years (average: 10 years) to evaluate whether their pretreatment serum lactate dehydrogenase (LDH) enzyme levels had a clinical value in predicting the course of the disease. The percentage of patients who had an elevated serum LDH at the time of diagnosis was significantly higher in those patients with metastatic disease than those who had localized disease (64% versus 33%, p < 0.0001), For those who presented with localized disease and had an increased serum LDH level, far more ultimately developed a relapse of disease (60% versus 38%, p < 0.0001) than those patients with a normal pre-treatment value. The prognostic significance of the serum LDH was more pronounced for the 247 patients treated with adjuvant chemotherapy (relapse rate of 72% versus 48%: p < 0.0002) than the 271 patients treated with neoadjuvant chemotherapy (relapse rate: 46% versus 28%, p < 0.005). Following treatment, serum LDH levels almost uniformly returned to normal and no correlation between postoperative levels and relapse of disease could be identified.

We have demonstrated that in patients with osteosarcoma of the extremities, pretreatment serum LDH levels have a definite prognostic value which should be considered when comparing the results achieved with different therapeutic protocols and in planning new randomized clinical trials.     

Gemcitabine for the Treatment of Patients with Osteosarcoma

Background: Patients with recurrent or refractory osteosarcoma are considered to have a very poor prognosis, and new regimens are needed to improve the prognosis in this setting. Gemcitabine, a nucleoside antimetabolite, is an analog of deoxycytidine which mainly inhibits DNA synthesis through interfering withDNA chain elongation and depleting deoxynucleotide stores, resulting in gemcitabine-induced cell death. Here we performed a systemic analysis to evaluate gemcitabine based chemotherapy as salvage treatment for patientswith recurrent or refractory osteosarcoma. Methods: Clinical studies evaluating the impact of gemcitabine based regimens on response and safety for patients with osteosarcoma were identified by using a predefined search strategy. Pooled response rates (RRs) of treatment were calculated. Results: In gemcitabine based regimens, 4 clinical studies which included 66 patients with recurrent or refractory osteosarcoma were considered eligible for inclusion. Systemic analysis suggested that, in all patients, pooled RR was 12.1% (8/66) in gemcitabinebased regimens. Major adverse effects were hematologic toxicity, including grade 3 or 4 anemia, leucopenia and thrombocytopenia in gemcitabine based treatment. No treatment related death occurred in gemcitabine based treatment. Conclusion: This systemic analysis suggests that gemcitabine based regimens are associated with mild activity with good tolerability in treating patients with recurrent or refractory osteosarcoma.     

Predictive Impact of Common Variations in DNA Repair Genes on Clinical Outcome of Osteosarcoma

We aimed to assess the role of XPG, XPC and MMS19L polymorphisms on response to chemotherapyin osteosarcomas, and the clinical outcomes. One hundred and eighty five osteosarcoma patients who werehistologically confirmed were enrolled in our study between January 2007 and December 2009. Genotyping ofXPG, XPC and MMS19L was performed in a 384-well plate format on the MassARRAY® platform. Individualswith XPG TT genotype and T allele were more likely to be better response to chemotherapy than CC genotype,with the OR (95% CI) of 4.17 (1.64-11.54) and 2.66 (1.39-5.11), respectively. Those carrying MMS19L TT genotypeand T allele showed better response to chemotherapy, with ORs (95% CI) of 4.8 (1.56-17.7) and 2.3 (1.22-4.36),respectively. Patients carrying TT genotype of XPG and MMS19L showed a significantly longer overall survivalthan CC genotype, with a 0.47 and 0.30-fold risk of death when compared with the wild-type of the gene. XPGand MMS19L are correlated with response to chemotherapy and prognosis of osteosarcoma, so that they couldbe used as predictive markers for prognosis.     

Survivin基因在骨肉瘤中的表达及其与临床耐药的相关性研究(英文)

目的探讨 Survivin 基因在骨肉瘤中的表达及其与临床耐药的相关性。方法采用免疫组织化学 S-P 法检测凋亡抑制基因 Survivin 在骨肉瘤、骨软骨瘤和正常骨组织中的表达及 P-糖蛋白在骨肉瘤中的表达。结果 Survivin 基因在骨肉瘤中的阳性表达率为65.71%,在骨软骨瘤和正常骨组织中未见表达;Survivin 阳性表达与患者的年龄、性别及肿瘤部位无关,与 Enneking 外科分期及 WHO 组织学分型有相关性;P-糖蛋白阳性表达率分别为45.7l%,Survivin 的表达与P-糖蛋白表达密切相关。结论 Survivin 在骨肉瘤中呈高表达,与临床耐药密切相关,有望成为骨肉瘤靶向治疗的一个新靶点。     

Survivin基因在骨肉瘤中的表达及其与临床耐药的相关性研究

目的探讨Survivin基因在骨肉瘤中的表达及其与临床耐药的相关性。方法采用免疫组织化学S-P法检测凋亡抑制基因Survivin在骨肉瘤、骨软骨瘤和正常骨组织中的表达及P-糖蛋白在骨肉瘤中的表达。结果Survivin基因在骨肉瘤中的阳性表达率为65.71%,在骨软骨瘤和正常骨组织中未见表达;Survivin阳性表达与患者的年龄、性别及肿瘤部位无关,与Enneking外科分期及WHO组织学分型有相关性;P一糖蛋白阳性表达率分别为45.71%,Survivin的表达与P-糖蛋白表达密切相关。结论Survivin在骨肉瘤中呈高表达,与临床耐药密切相关,有望成为骨肉瘤靶向治疗的一个新靶点。     

Predictors of Chemotherapy Induced Adverse Events in Pediatric Osteosarcoma Patients

Objective: To investigate the prevalence of chemotherapy-induced adverse events and the associated risk factors in pediatric patients with osteosarcoma. Methods: This retrospective cross-sectional study enrolled 90 pediatric osteosarcoma patients (with 1,017 chemotherapy cycles) treated at Srinagarind Medical Center, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand, between January 1, 2008 and December 31, 2018. The prevalence of major adverse events and a correlation between baseline characteristics and adverse events were analyzed using a generalized estimating equation model. Result: The prevalence of adverse events in 90 pediatric osteosarcoma patients (with 1,017 chemotherapy cycles) was determined as chemotherapy-induced nausea and vomiting (29.2%; n=296), hepatotoxicity (21.2%; n=215), anemia (70.69%; n=719), neutropenia (26.65%; n=271), and thrombocytopenia (13.65%; n=139). Factors associated with chemotherapy-induced hepatotoxicity included methotrexate dose ≥ 12 g/m2 (odds ratio [OR] 1.30; 95% confidence interval [CI] 1.22–1.39; P<0.001), plasma concentration of methotrexate at 72 hours >0.1 μM (OR 1.22; 95% CI 1.19–1.25; P<0.001), and pre-hydration rate ≤ 125 mL/m2/h (OR 1.10; 95% CI 1.07–1.12; P<0.001). Conclusion: Major adverse events are becoming more common in pediatric osteosarcoma patients, and risk factors include larger chemotherapy doses, higher plasma methotrexate concentrations, and a slower pre-hydration rate. The outcomes of the study could aid in the better treatment of toxicity in children with osteosarcoma.     

Investigating the Relationship between of Vascular Endothelial Growth Factor and HER-2neu in IHC Staining with Metastasis and Mortality in Patients with Osteosarcoma

Background: The expression of HER-2neu and vascular endothelial growth factor (VEGF) in patients with osteosarcoma may determine the response to treatment. These two factors are likely to be effective in cancer progression. This study aimed at investigating the prevalence of these two factors in the pathological samples. Methods: Pathological samples of patients with osteosarcoma collected at a cancer surgery center between 2017 and 2018 were evaluated, of which 37 samples were included. The samples were evaluated using the IHC technique by two pathologists. Results: 12 women and 25 men with an average age of 26.7 years were studied. 21 patients (56.8%) developed metastases from the beginning or during follow-up, whereas 16 patients (43.2%) have not yet developed metastases. Regarding HER-2neu, 21 patients (56.8%) scored 0, 9 patients (24.3%) scored 1, 3 patients (8.1%) scored +2, and 4 patients (10.8%) scored +3. The VEGF intensity scores of 0, 1+, 2+, +3, +4 and were found in 7 (18.9%),  2 (5.4%), 18 (48.6%), 8 (21.6%), and 2 (5.4%) patients, respectively.  The results of the study did not show a significant relationship between age, gender, metastasis, and positive expression rates of HER-2neu and VEGF. Conclusion: The high expression of VEGF (75.7%) in the studied samples should be considered and further studies on this biomarker in cases with osteosarcoma are recommended from different aspects. To achieve validated results and prove the results of this study, similar studies with a larger sample size should be performed, and using targeted therapy for angiogenesis in large scale trials should be considered.     

Preoperative Intra-Arterial Chemotherapy in Patients with Osteosarcoma: Case report of seven patients below the age of 18 years

Summary

In the period between 1987 and 1989, seven patients under 18 years of age with osteosarcoma of a lower limb were treated by preoperative intra-arterial chemotherapy. The age of the patients ranged from 12 to 17 years, the median age being 14.5 years. The polyethylene catheter was placed surgically into the lower epigastric artery. Two combinations of cytostatics were administered: adriamycin-bleomycin-cisplatin and high-dose methotrexate – vincristine – cisplatin. All patients, after two or three courses, underwent surgical resection of limb tumor. Tumor destruction ranged from 20 to 100%.

Four patients with necrosis from 80–100% remained free of disease from 18 to 30 months: two, having necrosis of 40% and 95% respectively, died. The youngest patient whose necrosis was as low as 20%, after completion of the systemic chemotherapy, developed local recurrence and pulmonary metastases.     

CXCL12 Expression is Predictive of a Shorter Time to Tumor Progression in Low-Grade Glioma: A Single-Institution Study in 50 Patients

Summary The clinical course of 50 patients with low-grade glioma (31 male, 19 female) undergoing surgery at a single Institution from 1992 to 1996 was analyzed in relationship with known prognostic factors as far as time to tumor progression (TTP) and survival time (ST) are concerned. Moreover, microvessel density (MVD) and expression of the angiogenesis-related chemokine CXCL12 were investigated in surgical specimens. Age at diagnosis ranged from 1 to 68 years (median 30). Histology revealed 11 fibrillary, 6 protoplasmatic, 5 gemistocytic astrocytoma, 18 oligoastrocytoma and 10 oligodendroglioma. Mean follow-up was 86 months. Four patients were lost to follow-up. Of the remaining 46, twenty-four have shown disease progression and 14 have died. Median overall survival was not achieved; an estimated 75% percentage of survivors was found at 78 months. Complete gross tumor removal was associated to a longer TTP (P = 0.04 logrank). Of the investigated immunohistochemical parameters, while MVD was not predictive of subsequent TTP, expression of CXCL12 was associated with a significantly shorter TTP (P = 0.01 logrank): this predictive value remained significant (P = 0.02) at multivariate analysis. The data suggest the possible prognostic value for CXCL-12 (an angiogenesis- and tumor-growth-related chemokine) on TTP in low-grade gliomas.     

Influence of Adriamycin Dose in the Outcome of Patients with Osteosarcoma Treated with Multidrug Neoadjuvant Chemotherapy: Results of Two Sequential Studies

Summary

The results of two sequential studies of neoadjuvant chemotherapy for osteosarcoma of the extremities performed at Rizzoli Institute between 1986 and 1991 in 228 patients are presented. In both studies preoperative chemotherapy consisted of two cycles of high dose methotrexate (HDMTX), cisplatinum (CDP) and adriamycin (ADM). Postoperatively the good responder patients were treated with the same drugs used before surgery while in the poor responder patients ifosfamide was added to these three drugs. The preoperative treatment was the same in both studies while after surgery in the second protocol either the cumulative dose of ADM (270 mg/m² instead of 360 mg/m²) or the single dose per cycle of this drug (60 mg/m² instead of 90 mg/m²) was reduced. These changes in the last protocol were done to reduce the cardiotoxicity of ADM that was high in the first study (2 deaths and 1 heart transplantation). Since in the last protocol – in comparison with the first protocol – after surgery chemotherapy was restarted earlier and ADM was administered not as a single drug but in combination with the CDP the dose intensity of ADM was unchanged while the dose intensity of MTX, CDP and ifosfamide was higher than in the first study. The preliminary results of the 84 patients treated in the second study show a 2-year disease free survival significantly lower than that achieved in the 144 patients treated in the first study (37/51 – 73% vs 123/144 – 85%: P < 0.008). In addition, even if in the last study there were no cases of clinical cardiotoxicity due to ADM, there was a significantly higher percentage of severe myelodepression that led to two deaths for infectious complications. These results suggest that in neoadjuvant treatment of osteosarcoma the total dose of ADM and/or the single dose per cycle of the same drug are an important determinant of outcome and that increasing the dose-intensity of less toxic but less active agents, MTX, CDP and ifosfamide, at the expense of the more active and more toxic agent, ADM, can lead to a poorer outcome without reducing toxicity.     

Correlation of Mast Cell Density,Tumor Angiogenesis,and Clinical Outcomes in Patients with Endometrioid Endometrial Cancer

Background: Tumor angiogenesis has been demonstrated in several kinds of neoplasms. There is evidence that mast cells can produce many different chemical mediators with angiogenic properties. Since their specific role in female genital tract cancer has not been well understood, this study was conducted to determine correlations between among mast cell density, tumor angiogenesis, and clinical outcomes in patients with endometrioid adenocarcinoma of endometrium. Methods: Histologically, four-micrometer-thick haematoxylin and eosin stained slides of hysterectomy specimens were evaluated. Microvessels were highlighted by CD31 immunostaining and mast cells were stained with 0.1% toluidine blue. All clinicopathological characteristics were reviewed to determine their possible correlation to microvessel density and number of mast cells. Results: A total of 46 patients who underwent a complete staging surgery were eligible for this study. The median age was 55 years (range, 32-70 years) and the median follow-up was 27.0 months (range 3.6-83.8). Microvessels appeared to correlate to some extent with parity and the mean count was likely to be higher in women with non-menopausal status (p=0.07), advanced FIGO stage (p=0.09), and lymph node metastasis (p=0.08). However, there was no significant correlation between microvessel counts, mast cell density, and disease recurrence. Conclusion: Our data suggest that the number of microvessel counts and mast cell density do not affect clinical progression or recurrence of endometrioid endometrial cancer.     

儿童骨肉瘤患者保留骨骺的灭活再植术

目的探讨在有效大剂量化疗前提下的保留骨骺灭活再植术的可行性。方法自1999年开始在MMIA方案(由甲氨喋呤、异环磷酰胺、阿霉素组成)的有效大剂量化疗保护下对2例儿童骨肉瘤患者行保留骨骺的灭活再植术。术前严格遵循新辅助化疗的原则,行2个疗程的MMIA化疗。化疗后患者疼痛消失,肿块缩小,AKP、LDH 明显下降,X线片及MR示左股骨下端病变局限,边缘清晰,硬化及骨化明显,骨骺未受侵蚀。术后继续应用2疗程化疗。结果2例患者术后切口均一期愈合。随访分别为3年和4年,无复发和转移,肢体功能恢复理想。结论保留骨骺的灭活再植术是治疗儿童骨肉瘤的一种新方法,其在不增加局部复发率的前提下避免术后双侧肢体不等长,并改善了术后患者的肢体功能。但严格掌握手术适应证是至关重要的,否则将导致灾难性后果。     

儿童骨肉瘤患者保留骨骺的灭活再植术(英文)

目的探讨在有效大剂量化疗前提下的保留骨骺灭活再植术的可行性。方法自1999年开始在 MMIA 方案(由甲氨喋呤、异环磷酰胺、阿霉素组成)的有效大剂量化疗保护下对2例儿童骨肉瘤患者行保留骨骺的灭活再植术。术前严格遵循新辅助化疗的原则,行2个疗程的 MMIA 化疗。化疗后患者疼痛消失,肿块缩小,AKP、LDH明显下降,X 线片及 MR 示左股骨下端病变局限,边缘清晰,硬化及骨化明显,骨骺未受侵蚀。术后继续应用2疗程化疗。结果 2例患者术后切口均一期愈合。随访分别为3年和4年,无复发和转移,肢体功能恢复理想。结论保留骨骺的灭活再植术是治疗儿童骨肉瘤的一种新方法,其在不增加局部复发率的前提下避免术后双侧肢体不等长,并改善了术后患者的肢体功能。但严格掌握手术适应证是至关重要的,否则将导致灾难性后果。     

Apurinic/apyrimidinic endonuclease 1 induced upregulation of fibroblast growth factor 2 and its receptor 3 induces angiogenesis in human osteosarcoma cells

Tumor angiogenesis contributes to inferior prognosis in osteosarcoma. Apurinic/apyrimidinic endonuclease 1 (APE1) and fibroblast growth factor 2 (FGF2) and its receptor 3 (FGFR3) signaling pathway plays an important role in the angiogenic process. In this study we observed that high expression of APE1, FGF2 and FGFR3, and microvessel density are positively correlated with poor prognosis of osteosarcoma patients. Furthermore, the Cox model showed that the tumor size, FGF2 and its receptor 3 (FGFR3), and microvessel density were adverse prognostic factors. Based on our clinical data, and the fact that APE1 is involved in tumor angiogenesis, we hypothesize that it is very likely that APE1 may indirectly promote angiogenesis by upregulating fibroblast FGF2 and FGFR3. Our preliminary data show small interfering RNA‐mediated silence of APE1 experiments, which further supports this hypothesis. APE1‐small interfering RNA significantly inhibited tumor angiogenesis by downregulating in vitro expression of FGF2 and FGFR3 in human umbilical vein endothelial cells in Matrigel tube formation assay, and further inhibited tumor growth in vivo in a mouse xenograft model. Thus, the proposed APE1‐FGF2 and FGFR3 pathway may provide a novel mechanism for regulation of FGF2 and FGFR3 by APE1 in tumor angiogenesis.     

A Retrospective Comparative Analysis of Outcomes and Prognostic Factors in Adult and Pediatric Patients with Osteosarcoma

Osteosarcoma is the most common primary bone malignancy in both children and adults. Despite introduction of intensive multimodal treatment with chemotherapy and surgery, outcomes are still poor, especially for patients with metastatic disease and adults. Hence, there is an ongoing need for better prognostic markers and outcome data to inform management decisions in both the adult and pediatric setting. Here, we retrospectively analyzed 112 patients with bone osteosarcoma treated at two large adult and pediatric tertiary academic centers between 1989 and 2019. Patients were divided into an adult (≥18 years) and pediatric (<18 years) cohort for comparison. Our aim was to evaluate predictors of outcomes in pediatric and adult patients, with a specific focus on the role of methotrexate when added to a combination of doxorubicin-cisplatin; the prognostic value of tumor necrosis after neoadjuvant chemotherapy; and outlining any differences in outcomes between adults and pediatric patients that could inform clinical management. Adult patients treated with methotrexate-doxorubicin-cisplatin and those treated with doxorubicin-cisplatin had similar 5-year PFS (26%, 95%CI: 45.5%–10% vs. 50%, 95%CI: 69.6%–26.2%, p = 0.1) and 5-year OS (63%, 95%CI: 82%–34%, vs. 78%, 95%CI: 90.6%–52.6%, p = 0.5). In the adult cohort, there was no difference between patients with ≥90% necrosis and <90% necrosis in either 5-year PFS (42%, 95%CI: 71.1%–11.3% vs. 38%, 95%CI: 57.7%–18.2%, p = 0.4) or 5-year OS (85%, 95%CI: 97.8%–33.4% vs. 56%, 95%CI: 76.8%–27.6%, p = 0.4). In the pediatric cohort, compared to patients with <90% necrosis, those with ≥90% necrosis had significantly better 5-year PFS (30%, 95%CI: 49.3%–14.1% vs. 55%, 95%CI: 73.9%–38.5%, p = 0.003) and 5-year OS (64%, 95%CI: 80.8%–41.1% vs. 78%, 95%CI: 92%–60.9%, p = 0.04). Adult and pediatric patients had similar 5-year OS (69%, 95%CI: 83.2%–49.8% vs. 73%, 95%CI: 83.2%–59.3%, p = 0.8) and 5-year PFS (37%, 95%CI: 52.4%–22.9% vs. 43%, 95%CI: 56.2%–30.4% p = 0.3) even though the proportion of patients with ≥90% necrosis after neoadjuvant chemotherapy was higher for children compared to adults (60.3% vs. 30%, OR: 3.54, 95%CI: 1.38–8.46, p = 0.006). In conclusion, in adult patients, the addition of methotrexate to doxorubicin and cisplatin did not correlate with a significant survival benefit, questioning the therapeutic value of methotrexate overall. Our study confirms the prognostic utility of percent tumor necrosis after neoadjuvant chemotherapy in pediatric patients but not in adult patients. Lastly, this is one of the few reported studies where patients with osteosarcoma younger and older than 18 years had similar PFS and OS.     

Association of Preoperative Radiation Effect with Tumor Angiogenesis and Vascular Endothelial Growth Factor in Oral Squamous Cell Carcinoma

This study examined the relationship between tumor angiogenesis and the radiation-induced response, evaluated based on pathological changes, in oral squamous cell carcinoma patients treated with preoperative radiation therapy. Forty-one cases of squamous cell carcinoma treated with preoperative radiation therapy were investigated. Tumor angiogenesis was assessed by scoring the intratumor microvessel density (IMVD). Expression of vascular endothelial growth factor (VEGF) was also evaluated before and after preoperative radiotherapy. There was no correlation between IMVD in the specimens before therapy and the pathological response to radiation therapy. However, radiation therapy decreased IMVD in the specimens after therapy. A significant association was observed between VEGF expression and resistance to radiation therapy: only 4 of the 21 patients whose tumors exhibited a high level (2+ or 3+) of VEGF staining experienced a major (3+ or 4+) pathological response to radiation therapy. Furthermore, an increasing level of VEGF expression after radiation therapy was observed in non-effective (0 to 2+) response cases. These results suggest that VEGF expression and the induction of this protein are related to radiosensitivity and could be used to predict the effects of preoperative radiation therapy on oral squamous cell carcinoma.