Objective To investigate the expression level and clinical significance of melanoma antigen gene A (MAGEA) in osteosarcoma patients. Methods Compare gene expression profiles in osteosarcoma cell lines and osteoblasts with gene microarrays. Validation of differentially expressed genes was carried out by real-time polymerase chain reaction analysis. Corresponding protein levels were measures by Western blot analysis in osteosarcoma cell lines and by immunohistochemistry in osteosarcoma tissues. The staining intensity of immuno-histochemistry was correlated with clinical outcome , and its prognostic significance was analyzed. Results Sev-eral genes belonging to MAGEA increased significantly in all osteosarcoma cell lines and tumor tissue , but not in normal osteoblast cell. Patients with MAGEA expression has higher risk of lung metastasis (relative risk 2.79, 95% confidence interval, 1.12-6.93; P = 0.028) and lower five-year survival rates (39.6% ± 8.4% vs. 80% ± 8.9%, P = 0.01) compared with patients without MAGEA expression. Conclusions The expression of MAGEA increased in osteosarcoma , which inversely correlating with outcome of osteosarcoma patients.

OBJECTIVETo detect the expression of SCUBE3 in human osteosarcoma cell lines and surgical specimens of osteosarcomas and investigate its association with the patients' prognosis.

METHODSThe expression of SCUBE3 protein was detected in 5 osteosarcoma cell lines using Western blotting. CCK8 assay was used to assess the effect of SCUBE3 suppression mediated by two specific small interfering RNAs (siRNAs) on the proliferation of U2OS osteosarcoma cells, and the cell cycle changes were detected using flow cytometry. Immunohistochemistry was performed to detect the expression of SCUBE3 in 60 osteosarcoma tissues, and Kaplan-Meier method was performed for survival analysis of the patients.

RESULTSCompared with osteoblast hFOB1.19 cells, the osteosarcoma cell lines all showed significantly higher expressions of SCUBE3. In U2OS cells, suppression of SCUBE3 expression by siRNA significantly inhibited the cell proliferation (P<0.05). Kaplan-Meier survival analysis indicated that patients with high SCUBE3 expression had worse prognosis than those with low SCUBE3 expression (P=0.036).

CONCLUSIONSCUBE3 is up-regulated in the 5 osteosarcoma cell lines and in primary osteosarcoma tissues to promote the proliferation of osteosarcoma cells. A high SCUBE3 expression in osteosarcoma tissues is associated with a poor prognosis of the patients, suggesting that SCUBE3 can serve as a potential therapeutic target for osteosarcoma.

Bone Neoplasms ; metabolism ; pathology ; Calcium-Binding Proteins ; metabolism ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Humans ; Immunohistochemistry ; Kaplan-Meier Estimate ; Osteosarcoma ; metabolism ; pathology ; Prognosis ; RNA, Small Interfering ; Up-Regulation

OBJECTIVE： To study lung tissue injury induced by long-term aerosol inhalation of 4 kinds of non-aerosol drugs in healthy SD rats， and to evaluate the safety of aerosol inhalation of non-aerosol drugs. METHODS： Totally 40 healthy SD rats （♂） were randomly divided into 8 group， i.e. blank control group， normal saline group （solvent control）， budesonide group （non-aerosol drug control， 0.1 g/L） ，silicon dioxide group （lung injury drug control， 40 g/L）and 4 kinds of non-aerosol drugs [Dingchuan decoction group （15 g/mL， calculated by crude drug）， cefatriaxone group （200 g/L）， Qingkailing group （stoste） and Tangreqing group （stoste）]， with 5 rats in each group. Except that blank control group didn’t received any treatment， other groups received aerosol inhalation， 10 mL， twice a day， for consecutive 56 days. After medication， the number of white blood cells in peripheral blood were counted and classified， and the number of white blood cells in bronchus alveolar lavage fluid were counted. The pathological changes of lung tissue were observed by HE staining and the number of dust cells was counted. The expression of leukocyte differentiation antigen 163 （CD163） in lung tissue were determined by immunohistochemical method. RESULTS： The white blood cells in peripheral blood mainly included lymphocyte and neutrophil， of which lymphocyte is the main one. Compared with blank control group， there was no statistical significance in the number of white blood cells， lymphocyte or neutrophil in peripheral blood， the number of white blood cells in alveolar lavage fluid or the number of dust cells in lung tissue of rats in normal saline group （P＞0.05）； the structures of bronchus and lung tissue were intact， and the expression of CD163 was negative. Compared with normal saline group， there was no statistical significance in the above indexed of rats in budesonide group（P＞0.05）， the structures of bronchus and lung tissue were intact， and the expression of CD163 was negative， while the number of white blood cells， lymphocyte or neutrophil in peripheral blood， the number of white blood cells in alveolar lavage fluid or the number of dust cells in lung tissue of rats in other 5 groups were all increased significantly （P＜0.05）； alveolar wall thickening and alveolar interstitial edema occurred in different degrees in lung tissue. The expression of CD163 was positive or strongly positive. CONCLUSIONS： The long-term aerosol inhalation of 4 kinds of non-aerosol drugs can induce pathological changes of lung tissue and increase the number of inflammatory cell and dust cell in alveolin in healthy SD rats.