As a highly aggressive bone malignancy, osteosarcoma poses a significant therapeutic challenge, especially in the setting of metastasis or recurrence. This study aimed to investigate the potential of CD8-Tex cell-associated genes as prognostic biomarkers to reveal the immunogenomic profile of osteosarcoma and guide therapeutic decisions. mRNA expression data and clinical details of osteosarcoma patients were obtained from the TCGA database (TARGET-OS dataset). The GSE21257 dataset (from the GEO database) was used as an external validation set to provide additional information on osteosarcoma specimens. 84 samples from the TARGET-OS dataset were used as the training set, and 53 samples from the GSE21257 dataset served as the external validation cohort. Univariate Cox regression analysis was utilized to identify CD8 Tex cell genes associated with prognosis. The LASSO algorithm was performed for 1000 iterations to select the best subset to form the CD8 Tex cell gene signature (TRS). Final genes were identified using the multivariate Cox regression model of the LASSO algorithm. Risk scores were calculated to categorize patients into high- and low-risk groups, and clinical differences were explored by Kaplan–Meier survival analysis to assess model performance. Prediction maps were constructed to estimate 1-, 3-, and 5 year survival rates for osteosarcoma patients, including risk scores for CD8 Texcell gene markers and clinicopathologic factors. The ssGSEA algorithm was used to assess the differences in immune function between TRS-defined high- and low-risk groups. TME and immune cell infiltration were further assessed using the ESTIMATE and CIBERSORT algorithms. To explore the relationship between immune checkpoint gene expression levels and the two risk-defined groups. A CD8 Tex cell-associated gene signature was extracted from the TISCH database and prognostic markers including two genes were developed. The high-risk group showed lower survival, and model performance was validated by ROC curves and C-index. Predictive plots were constructed to demonstrate survival estimates, combining CD8 Tex cell gene markers and clinical factors. This study provides valuable insights into the molecular and immune characteristics of osteosarcoma and offers potential avenues for advances in therapeutic approaches.

目的 探讨β-榄香烯在骨肉瘤中诱导自噬发生的信号通路及其对肿瘤细胞增殖及凋亡的影响.方法 以不同浓度(0、10、20、50、100、150μg/ml)的β-榄香烯处理人骨肉瘤143B细胞24、48 h后,应用CCK-8法检测细胞存活率;用0、20、50μg/mlβ-榄香烯处理143B细胞48 h后,应用流式细胞仪使用AnnexinⅤ-FITC/PI双染法检测细胞凋亡率,并通过荧光显微镜及蛋白印迹法(Western blotting)检测细胞自噬相关蛋白微管相关蛋白1轻链3(LC3B)的分布及表达,Western blotting检测143B细胞中丝氨酸/苏氨酸激酶(AKT)、磷酸化丝氨酸/苏氨酸激酶(p-AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)及磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)蛋白表达水平.结果 β-榄香烯呈浓度依赖性地显著抑制骨肉瘤143B细胞的增殖,其对143B细胞24 h和48 h时的半数抑制浓度(IC50)分别为58.89μg/ml和18.48μg/ml;143B细胞经20、50μg/mlβ-榄香烯处理48 h后,细胞凋亡率分别为(15.43±0.99)％和(19.21±0.82)％,明显高于0μg/mlβ-榄香烯组(6.87±1.02)％;且其细胞Cleaved caspase-3蛋白表达水平亦明显高于0μg/mlβ-榄香烯组,差异均有统计学意义(P<0.05);经20、50μg/mlβ-榄香烯处理骨肉瘤143B细胞48 h后,细胞质内的自噬体数量明显增多,其LC3-Ⅱ/LC3-Ⅰ表达水平较0μg/mlβ-榄香烯组均明显增高,差异均有统计学意义(P<0.05),β-榄香烯可以抑制骨肉瘤细胞p-AKT和p-mTOR的表达,且呈时间及剂量依赖性.结论 β-榄香烯可以诱导人骨肉瘤143B细胞增殖抑制及凋亡,还可以通过抑制AKT/mTOR通路激活细胞自噬.

Osteosarcoma (OS) is the most common primary malignant bone tumor in pediatric and adolescent patients. The calcyclin-binding protein/Siah-1-interacting protein (CacyBP/SIP) performs an essential function in cell proliferation and apoptosis. The present study investigated the effect of CacyBP/SIP in OS cell proliferation and apoptosis. CacyBP/SIP mRNA expression levels were evaluated in four OS cell lines by quantitative PCR. CacyBP/SIP expression was downregulated in Saos-2 cells using a lentivirus transfection system and the transfection efficiency was analyzed. The effects of CacyBP/SIP downregulation on Saos-2 cell proliferation and colony-formation ability were evaluated by MTT and colony-formation assays. The effect of CacyBP/SIP knockdown on Saos-2 cell cycle and apoptosis was analyzed by flow cytometry cell sorting. The Cancer Genome Atlas (TCGA) data was analyzed for validation. Human OS cell lines Saos-2, MG-63, HOS and U20S expressed CacyBP/SIP mRNA. CacyBP/SIP knockdown significantly inhibited cell proliferation and colony-formation ability. G1/S phase arrest was induced by CacyBP/SIP downregulation, which also resulted in the downregulation of CDK and cyclins and the upregulation of p21. In addition, CacyBP/SIP downregulation induced Saos-2 cell apoptosis mediated by Bax and Bcl-2. High expression of CacyBP/SIP was significantly associated with poor prognosis in TCGA sarcoma database. Thus, CacyBP/SIP performs important functions in the proliferation and apoptosis of human OS cells.

Objective This study was performed to explore the relationship between various clinical factors and the prognosis of limb osteosarcoma. Methods We retrospectively analyzed the clinical data of 336 patients with limb osteosarcoma treated from June 2000 to August 2016 at 7 Chinese cancer centers. Data on the patients’ clinical condition, treatment method, complications, recurrences, metastasis, and prognosis were collected and analyzed. Kaplan–Meier analysis and Cox regression models were used to analyze the data. Results The patients comprised 204 males and 132 females ranging in age from 6 to 74 years (average, 21.1 years). The overall 3- and 5-year survival rates were 65.0% and 55.0%, respectively. The 5-year overall survival rate was 64.0% with standard chemotherapy and 45.6% with non-standard chemotherapy. Cox regression analysis demonstrated that standard chemotherapy, surgery, recurrence, and metastasis were independent factors associated with the prognosis of limb osteosarcoma. Conclusion The survival of patients with limb osteosarcoma can be significantly improved by combining standard chemotherapy and surgery. The overall survival rate can also be improved by adding methotrexate to doxorubicin–cisplatin–ifosfamide triple chemotherapy.

Objective The purpose of this retrospective study was to evaluate the clinical and oncological results of combination treatment of short‐term preoperative denosumab (the receptor activator of nuclear factor kappa‐B ligand inhibitor) with surgery in unresectable or recurrent cases of giant cell tumor of the bone (GCTB). Methods Between 2016 and 2018, 11 eligible patients (1 man, 10 women, mean age 38.1 years) with grade 3 GCTB were treated with a combination of short‐term (six doses) preoperative denosumab and surgery in a single institution. The clinical, radiological, and pathological alteration after the denosumab treatment were compared. The oncological results of the combination therapy were also recorded. Meanwhile, adverse effects or complications of denosumab, if any, were reported. Results The median follow‐up time after surgical procedure was 30 months (range 13–45 months). After 3–4 denosumab injections, pain relief was observed in all patients. In two spine patients, the neurological status improved after four doses of treatment. Intraoperatively, the margin of the tumor became clear and the intensity of the tumor increased while the blood supply around and within the lesion decreased. Within the lesion, the typically soft and loose tissue were replaced by the tough and dense fibro‐osseous tissue. The mean diameter of the lesion before and after treatment was 61.55 ± 22.49 mm and 51.81 ± 21.12 mm, respectively, and the T‐score was 1.02 (P = 0.32). Variable calcification was observed at the periphery and within the lesion. A total of three patients experienced local recurrence in this study. In the resection group, only one extremity patient had soft tissue recurrence that was treated with en‐bloc excision. In the curettage group, two of three sacral tumor patients had local occurrence. Both refused re‐operation and restarted the monthly denosumab injection thereafter, and the lesions remained stable at the final follow up. Finally, no adverse effects or complications related to denosumab treatment were found. Conclusion For the unresectable or recurrent GCTB cases, short‐term (six doses) preoperative use of denosumab improved clinical symptoms, decreased the tumor size, and increased the tumor density. The changes in tumors, in turn, simplified the tumor removal manipulation and, subsequently, decreased the local recurrence for the resection surgery. For the curettage, the denosumab‐induced changes had mixed impacts, and shorter term (fewer than six doses) usage may be more appropriate. Our six‐dose regime was deemed safe, while the safety of long‐term use remains unknown.

Di (2-ethylhexyl) phthalate (DEHP), which is widely used in polyvinyl chloride materials, can be easily accumulated into human body. Lung cancer cells can be directly exposed to DEHP via inhalation, however, the effects and related mechanisms of DEHP on the progression of non small cell lung cancer (NSCLC) were not illustrated. Our present study revealed that DEHP less than 10-4M had no significant effect on the proliferation of A549 or H1299 cells, while nanomolar DEHP can trigger the migration and invasion of NSCLC cells. DEHP treatment also increased the expression of interleukin-6 (IL-6) and IL-8. Silencing of IL-6, while not IL-8, can attenuate DEHP induced migration and invasion of NSCLC cells. This was confirmed by result that neutralization antibody of IL-6, while not anti-IL-8, attenuated DEHP induced invasion of A549 cells. The inhibitor of NF-κB, while not ERK1/2 or Akt, abolished DEHP induced up regulation of IL-6 and invasion of NSCLC cells. DEHP treatment can increase the phosphorylation, nuclear localization, and transcriptional activities of p65, one of the major parts of NF-κB complex. Collectively, our study revealed that DEHP can stimulate NSCLC migration and invasion via NF-κB mediated up regulation of IL-6.