摘要
目的 :观察缺氧条件下骨肉瘤SaOS 2细胞系缺氧诱导因子HIF 1α和血管生长因子VEGF表达的变化。方法 :建立骨肉瘤细胞体外缺氧培养模型 ,观察缺氧培养不同时相 (8、16、2 4h)HIF 1α和VEGF的表达。半定量RT PCR方法检测HIF 1α和VEGFmRNA的转录水平 ,免疫组化 (SP法 )和免疫印迹 (Westernblot)检测HIF 1α蛋白表达情况 ,免疫组化和ELISA法检测细胞和培养上清液中VEGF蛋白表达。结果 :缺氧处理后 ,HIF 1αmR NA转录水平没有明显改变 ,蛋白表达随缺氧时间延长而升高。VEGFmRNA以及蛋白的表达在缺氧条件下均显著增强。结论 :骨肉瘤细胞系SaOS 2在缺氧条件下 ,缺氧诱导因子HIF 1α和血管生长因子VEGF蛋白表达上调。
Purpose:To investigate the change of hypoxia in ducible factor 1α(HIF-1α) and vascular endothelial growth factor (VEGF) under hypoxic environment in vitro in osteosarcoma cell line SaOS-2. Methods:The hypoxia culture model was established by a hypoxia chamber. The gene productions of HIF-1α and VEGF were observed at different hy poxia culture phase. The mRNA expression of HIF-1α and VEGF was shown by the s emi-quantitative reverse transcription PCR(RT-PCR). The HIF-1α protein was t ested using immunohistochemical staining and Western Blot analysis. The VEGF pro tein in SaOS-2 and supernatants was tested using immunohistochemical staining a nd enzyme-linked immunosorbent assay (ELISA). Results:After the hypoxia treatment,the mRNA level of HIF-1α was not changed significantly ,however, the protein expression was increased rem arkably with correspondence to the hypoxia time. VEGF expression was up-regula ted under hypoxia both in mRNA level and in protein level. Conclusions:HIF-1α and VEGF genes could be up-regulated in o steosarcoma cell line SaOS-2 by hypoxia culture in vitro. These mechanism m ay be involved in the progress of tumor angiogenesis.
出处
《中国癌症杂志》
CAS
CSCD
2004年第6期501-504,共4页
China Oncology
关键词
骨肉瘤
缺氧
血管生长因子
缺氧诱导因子
osteosarcoma
hypoxia
vascular endothelial g rowth factor
hypoxia inducible factor 1α
