缺氧对骨肉瘤细胞系MG63 HIF1α、p53、bcl2及细胞凋亡影.docVIP

缺氧对骨肉瘤细胞系MG63 HIF1α、p53、bcl2及细胞凋亡影 　作者：蔡文涛 陈安民 郭风劲 管频 【摘要】 目的了解缺氧环境对骨肉瘤MG63细胞缺氧诱导因子HIF1α、p53、bcl2的表达及细胞凋亡的 影响 。 方法 建立骨肉瘤细胞体外缺氧模型，观察不同缺氧时间段(8、16、24h)HIF1α、p53、bcl2的表达和细胞凋亡的情况。半定量RTPCR方法检测HIF1α、p53、bcl2的表达水平；免疫组化(SP法)和免疫印迹(Western Blot)检测HIF1α、p53、bcl2蛋白表达情况；流式细胞仪检测细胞凋亡率。结果缺氧处理后，HIF1α转录水平未见明显改变，蛋白表达水平随缺氧时间延长明显增强；而p53、bcl2 mRNA及蛋白表达水平均显著增强，两者间存在一定的相关性；细胞凋亡率却未见明显增加。结论在缺氧条件下，不能通过以HIF1α为中介的p53依赖途径来诱导骨肉瘤MG63细胞的凋亡，其机制可能与缺氧诱导野生型p53的突变或缺失使HIF1α和bcl2的过表达有关。 【关键词】 骨肉瘤；缺氧诱导因子1α；p53；bcl2；细胞凋亡 论文代写 　　Effect of Hypoxia on the Expression of Hypoxia Inducible Factor 1α、p53、bcl2 and Apoptosis in Osteosarcoma Cell Line MG63 　　Abstract：ObjectiveTo investigate the effect of hypoxia on the expression of ypoxia inducible factor 1α(HIF1α)、p53、bcl2 and apoptosis in osteosarcoma cell line MG63 under hypoxia environment.Methods The hypoxia culture model was established by a hypoxia incubator.The gene productions of HIF1α、p53、bcl2 and the changes of the apoptosis were observed at different hypoxia culture phase.The semiquantitative reverse transcriotion PCR(RTPCR) was used to test the mRNA expression of HIF1α、p53 and bcl2.The protEin level of HIF1α、p53 and bcl2 was observed by immunohistochemical staining and western blot analysis.The changes of apoptosis were analyzed using flow cytometry.ResultsAfter the hypoxia treatment,the mRNA level of HIF1α was not changed significantly,however,the protEIn expression of HIF1α was increased remarkably with correspondence to the hypoxia time.But the expression of p53 and bcl2 were upregulated in mRNA and protein level.Besides,the bcl2 activity was markedly associated with the level of HIF1α. However ,the apoptosis rate was not increased markedly.ConclusionIn hypoxia environment,the apoptosis of osteosarcoma cell line MG63 could not induce by the p53 avenue which depended on HIF1α.The mechanism is possibly related with the mutation of p53 that increases the expression of HIF1α and bcl2. 　　Key