目的 探讨骨肉瘤（OS）和软组织肉瘤（STS）的关键核心基因（Hub基因）及其潜在作用,为肿瘤诊断和预后提供新依据。方法 从基因表达综合数据库（GEO）获得OS数据集GSE16088及STS数据集GSE21122,采用GEO2R在线工具筛选GSE16088和GSE21122数据集的差异表达基因（DEGs）。通过韦恩图获得2个数据集共同DEGs。选取2个数据集中差异表达最显著的上调和下调基因各20个,分别绘制聚类热图。通过使用注释、可视化和综合发现数据库（DAVID）对2个数据集的共同DEGs进行功能（GO）和通路（KEGG）富集分析。构建蛋白互作网络并使用最大中心度（MCC）算法筛选排名最前的10个基因作为潜在的关键Hub基因。采用受试者工作特征（ROC）曲线探讨关键Hub基因对肉瘤患者的诊断价值。通过Kaplan-Meier Plotter进行生存期分析。通过实时荧光定量PCR技术对得分靠前的5个Hub基因进行验证。结果 GSE16088数据集筛选出5 210个DEGs,其中上调和下调的DEGs分别为1 028、4 182个;GSE21122数据集共筛选出1 224个DEGs,其中上调和下调的DEGs分别为451、773个;2个数据集共获得498个共同DEGs。共同DEGs参与到多个生物学过程和信号通路。基于PPI网络和MCC算法最终获得10个关键Hub基因,ROC曲线验证结果符合预期,且生存期分析10个关键Hub基因与肉瘤预后显著相关（P<0.05）。Hub基因在mRNA表达水平和生物信息学分析结果一致（P<0.05）。结论 10个关键Hub基因可用于肉瘤的诊断和预后,为后续免疫治疗提供新视野。

Objective To explore the Hub genes of osteosarcoma（OS）and soft tissue sarcoma（STS）and their potential roles,and to provide evidence for tumor diagnosis and prognosis．Methods The GSE16088 dataset and the GSE21122 dataset were screened in the Gene Expression Omnibus database of the National Center for Biotechnology Information in the United States．The online editing tool GEO2R was used to screen the DEGs of the GSE16088 dataset and the GSE21122 dataset and the Veen map was drawn to find the common DEGs of the GSE16088 dataset and the GSE21122 dataset．20 up-regulated and 20 down-regulated genes with the most significant differential expression were selected from 2 datasets,and heatmaps were drawn for each．The Database for Annotation,Visualization and Integrated Discovery was used to perform GO function enrichment analysis and KEGG pathway enrichment analysis on DEGs of GSE16088 dataset and GSE21122 dataset．PPI network of DEGs was constructed by STRING．PPI sub-modules and Hub genes with high connectivity were screened．Maximal clique centrality（MCC）score was used to select the Hub genes in the protein interaction network．The predictive value of 10 Hub genes in sarcoma patients was analyzed by receiver operating characteristic（ROC）curve．Survival analysis was performed by means of the Kaplan-Meier Plotter．The top five core genes were verified by real-time fluorescence quantitative PCR．Results A total of 5 210 genes were screened in GSE16088 dataset,including 1 028 and 4 182 genes with upregulated and downregulated expression．A total of 1 224 genes were selected from the GSE21122 dataset,including 451 and 773 genes with upregulated and downregulated expression．The cluster heatmap was used to show the top 20 DEGs with high and low expression in GSE16088 and GSE21122 datasets．By differential analysis of gene expression between the two datasets,498 co-DEGs were obtained．GO and KEGG enrichment showed that common DEGs were associated．Ten Hub genes were obtained by PPI and MCC algorithm,the ROC curve verification results were as expected．Survival analysis showed that 10 Hub genes were significantly associated with the prognosis of sarcoma（P<0.05）．The mRNA expression level of Hub genes was the same as the results of bioinformatics analysis（P<0.05）．Conclusions The 10 Hub genes can be used for the diagnosis and prognosis of osteosarcoma,and provide a new vision for subsequent immunotherapy．

目的 体外细胞实验检测吴茱萸碱对骨肉瘤HOS细胞株细胞周期及体外增殖能力的影响。方法 通过利用浓度为0、3、6、12 μmol/L吴茱萸碱处理骨肉瘤HOS细胞48 h后,Hoechst-33258荧光染色观察不同浓度吴茱萸碱处理后HOS细胞核的形态学变化。利用流式细胞术检测3 μmol/L的吴茱萸碱处理后骨肉瘤HOS细胞的细胞周期分布变化。结果 3、6、12 μmol/L的骨肉瘤吴茱萸碱处理细胞,细胞呈现凋亡核碎裂等典型变化,而且随药物剂量增加而更趋明显。呈剂量依赖性抑制其体外增殖能力。3 μmol/L吴茱萸碱处理骨肉瘤HOS细胞0、24、48、72 h,各组细胞周期变化如下:G0/G1期:对照组(51.12±2.13)%、24 h(19.17±1.02)%、48 h(16.94±1.67)%、72 h(11.05±1.25)%;S期:对照组(32.92±0.73)%、24 h(31.00±1.42)%、48 h(32.38±3.03)%、72 h(29.18±2.87)%;G2/M期:对照组(16.01±2.26)%、24 h(49.82±0.62)%、48 h(50.6767±2.80)%、72 h(59.56±1.97)%。结论 吴茱萸碱可诱导人骨肉瘤HOS细胞发生G2/M期阻滞,而S期变化不明显。说明吴茱萸碱可以抑制骨肉瘤细胞的增殖能力,并阻滞细胞周期于G2/M期。

Objective Using transcriptome sequencing and in vitro cell assay to detect the effect of evodiamine on cell cycle and proliferation in osteosarcoma HOS cell line. Methods HOS cells were treated with evodiamine at 0, 3, 6, and 12 μmol/L for 48 hours, Hoechst-33258 fluorescence staining was used to observe the morphological changes of HOS nuclei after treatment with different concentrations of evodiamine.The cell cycle distribution of HOS cells treated with 3 μmol/L evodiamine was detected by flow cytometry. Results 3,6,12 μmol/L osteosarcoma treated with evodiamine, the cells showed typical changes such as apoptotic nuclear fragmentation, and it became more obvious with the increase of drug dosage. Inhibition of proliferation in vitro in a dose-dependent manner.HOS cells were treated with 3 μmol/L evodiamine for 0, 24, 48, 72 h. The cell cycle changes of each group were as follows: G0/G1 phase: control group(51.12±2.13)%, 24 h(19.17±1.02)%, 48 h(16.94±1.67) %, 72 h(11.05±1.25)%;S phase: control group(32.92±0.73)%, 24 h(31.00±1.42)%, 48 h(32.38±3.03)%, 72 h(29.18±2.87)%;G2/M period: control group(16.01±2.26)%, 24 h(49.82±0.62)%, 48 h(50.6767±2.80)%, 72 h(59.56±1.97)%. Conclusion Analysis of the above results revealed that evodiamine can induce G2/M phase arrest in human osteosarcoma HOS cells, but the S phase changes are not obvious. It indicated that evodiamine would inhibit the proliferation of osteosarcoma cells and block the cell cycle in G2/M phase.

目的 研究新辅助化疗联合保肢手术治疗骨肉瘤的临床效果。方法 根据既往治疗骨肉瘤方法的不同,将56例患者分为对传统保肢组（A组）和现代保肢组（B组）,各28例。A组用采用传统保肢方案,即保肢手术+术后化疗方案进行;B组采用现代保肢方案,即新辅助化疗+保肢手术+术后化疗方案进行。化疗方案均采用CTX + VCR +MTX+ ADM方案,比较两组转移/复发率、术后1 年、2年及 3 年生存率、肢体功能、临床疗效情况。结果 比较两组的转移/复发率及3年后的死亡率,B组低于A组(P<0.05);肢体功能优良率及临床疗效,B组高于A组(P<0.05)。结论 新辅助化疗联合保肢手术能够降低骨肉瘤患者转移/复发率、死亡率,改善肢体功能,提高临床疗效。

Objective To study the clinical effect of neoadjuvant chemotherapy combined with limb salvage surgery for osteosarcoma. Methods Based on the previous methods of treating osteosarcoma, 56 patients were divided into the traditional limb salvage group (A group) and the modern limb salvage group (B group), 28 cases for each. The traditional limb salvage surgery + postoperative chemotherapy regimen were used in group A, and modern limb salvage regimen was performed in group B, ie neoadjuvant chemotherapy + limb salvage surgery + postoperative chemotherapy. The CTX + VCR +MTX+ ADM protocol was used in the chemotherapy regimens. The metastasis/recurrence rate, the 1-year, 2-year and 3-year survival rates, limb function, and clinical efficacy were compared between the two groups. Results The metastasis/recurrence rate and the mortality rate after 3 years in the two groups were compared. The B group was lower than that of the group A (P<0.05). The excellent rate of limb function and clinical efficacy were higher in the B group than that of in the group A (P<0.05). Conclusion Neoadjuvant chemotherapy combined with limb salvage surgery may reduce the metastasis/recurrence rate and mortality of osteosarcoma patients, improve limb function and increase clinical efficacy.