ERK1/2 pathway mediates the protective effect of hydrogen sulfide against cisplatin-induced injury in human marrow mesenchymal stem cells / 中华骨科杂志

Objective To explore the protection and mechanism of hydrogen sulfide (H2S) preconditioning against injury induced by cisplatin in human marrow mesenchymal stem cells (HMMSCs).Methods HMMSCs were treated by cisplatin at 0,5,10,20,40,60,80 mg/L concentrations for 24 h respectively and were exposed to cisplatin at 20 mg/L concentrations for 0,6,12,24,36,48 h respectively.HMMSCs were pretreated with NaHS at 0,0.4,0.6,0.8,1.0 mmol/L respectively for 30 min before exposed to cisplatin at 20 mg/L concentrations for 24 h.HMMSCs were treated by U0126 or combined with human epidermal growth factor (HEGF) together for 30 min before they were preconditioned with NaHS for 30 min.The cell survival rate,cell apoptosis rate,the expression of p-ERK1/2 and t-ERK1/2 were recorded.Results The cell survival rate decreased to 71.72％±2.72％,59.41％±5.44％,50.37％±4.55％,38.97％±2.92％,30.11％±4.64％ and 21.71％±5.35％ respectively after cells were treated with cisplatin at 5,10,20,40,60,80 mg/L concentrations respectively,and the differences were statistically significant compared with 0 mg/L group.The cell viability fell to 70.30％±6.20％,61.63％±2.70％,51.29％±3.13％,38.72％±3.66％ and 27.57％±2.32％ after HMMSCs were treated with cisplatin at 20 mg/L for 6,12,24,36,48 h respectively,and the differences were statistically significant compared with 0 h group.The cell viability increased to 65.99％±2.67％,72.93％±5.44％,75.10％±4.71％ and 76.56％± 5.25％ when HMMSCs got pretreatment of NaHS,and the differences had statistical significance compared with cisplatin group.The cell apoptotic rate decreased from 35.29％±2.77％ to 18.62％±0.97％ when HMMSCs were pretreated with NaHS at 0.6 mmo/L.Treatment of HMMSCs with cisplatin at 20 mg/L for 24 h reduced p-ERK1/2 expression.The pretreatment of NaHS could inhibit the inhibitory action to the expression of p-ERK1/2 induced by cisplatin.Pretreatment with U0126 or HEGF inhibited or promoted the protection and the upregulated expression ofp-ERK1/2 caused by NaHS pretreatment.Conclusion The preconditioning of H2S can protect cell damage caused by cisplatin via activating the ERK1/2 pathway of HMMSCs.

Expression of chemokine CXCL14 in primary osteosarcoma and its association with prognosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expression of CXCL14 in human osteosarcoma cell lines and tissues and investigate its association with the prognosis of the patients.</p><p><b>METHODS</b>RT-PCR, enzyme-linked immunosorbent assay (ELISA) and real-time PCR were used to detect the expression of CXCL14 in 4 osteosarcoma cell lines and in 40 pairs of osteosarcoma tissues and adjacent muscular tissues. CCK8 assay and colony formation assay was used to assess the effect of CXCL14 suppression mediated by two specific siRNAs on the proliferation of U2OS osteosarcoma cells. Immunohistochemistry was performed to detect the expression of CXCL14 in 58 osteosarcoma tissues, and Kaplan-Meier method and log-rank test were performed for survival analysis of the patients.</p><p><b>RESULTS</b>Significant up-regulation of CXCL14 expression was found in the osteosarcoma cell lines and in osteosarcoma tissues compared with the adjacent muscles (P<0.01). In U2OS cell, suppression of CXCL14 expression by siRNA significantly inhibited the cell proliferation (P<0.01) and colony formation rate (P<0.05). Kaplan-Meier survival analysis indicated that patients with high CXCL14 expression had worse prognosis than those with low CXCL14 expression (P=0.02).</p><p><b>CONCLUSION</b>CXCL14 is up-regulated in both osteosarcoma cell lines and primary osteosarcoma tissues to promote the proliferation of osteosarcoma cells. A high CXCL14 expression in osteosarcoma tissues is associated with a poor prognosis, suggesting the that CXCL14 serve as a potential therapeutic target for osteosarcoma.</p>

Survival analysis of 311 patients with osteosarcoma of the extremities treated in a single institute / 中华骨科杂志

Objective To investigate treatment and prognosis of patients with osteosarcoma of the extremities.Methods A total of 311 patients with osteosarcoma of the extremities,who had undergone treatment in our institute from 1998 to 2008,were enrolled in this retrospective study.Kaplan-Meier survival curve and Cox regression model were used to analyze the correlation between survival rate and variables including patients' demographics,chemotherapy,surgery,complications,and tumor metastasis.Results Among 311 patients,there were 206 males and 105 females,aged from 5 to 56 years (average,18.6 years).A total of 282 patients underwent aggressive or radical surgery,including 149 cases of limb salvage surgery and 133 cases of amputation surgery.One hundred and five patients underwent standard chemotherapy and 206 patients underwent non-standard chemotherapy.The 5-year survival rate was 57.4％ in patients treated with standard chemotherapy,36.3％ in patients treated with non-standard chemotherapy,16.8％ in patients with lung metastasis,50.7％ in patients without lung metastasis,56.6％ in patients who underwent limb salvage surgery,31.8％ in patients who underwent amputation surgery,44.6％ in patients with Enneking stage Ⅱ B and 33.1％ in patients with Enneking stage Ⅲ.For patients treated by amputation surgery,because non-standard chemotherapy which was performed in most of them and other confounding factors,the 5-year survival rate of them was lower.The Cox regression analysis showed that lung metastasis and non-standard chemotherapy were associated with inferior outcomes.Conclusion Neoadjuvant chemotherapy combined with aggressive or radical surgery could cure about 60％ of patients with osteosarcoma of the extremities.Lung metastasis and non-standard chemotherapy are risk factors that severely affect prognosis.