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作 者:金问森[1] 孔肇路[1] 沈芝芬[1] 童顺高[1] 季华均[1] 金一尊[1]
机构地区:[1]复旦大学放射医学研究所第八研究室,上海200032
出 处:《中华放射医学与防护杂志》2008年第1期1-4,共4页Chinese Journal of Radiological Medicine and Protection
基 金:国家高技术研究发展计划“863”项目(2002AA2Z3104);国家自然科学基金资助项目(30500143,30400089)
摘 要:目的研究辐射对肝癌细胞内乏氧诱导因子1α(HIF-1α)的调节作用以及HIF-1α的变化对肝癌细胞存活的影响。方法采用CoCl2化学模拟乏氧预处理人肝癌HepG2细胞,经不同剂量的γ射线照射后,通过细胞集落形成实验,比较有氧及化学模拟乏氧条件下细胞存活水平的差异。同时采用RT-PCR和Western blot方法检测照射后细胞内HIF-1α在转录和翻译水平上的变化。结果在化学模拟乏氧条件下,细胞存活分数(SF)显著增加,与有氧对照组比较差异有统计学意义,且两种不同作用条件下的SF比值(SFco/SFo2)随照射剂量逐渐增加,同时,经照射后细胞内的HIF-1α表达升高,并呈剂量依赖模式,HIF-1α的表达与照射后的细胞存活水平具有较强的关联性。结论辐射可诱导肝癌细胞内HIF-1α的表达升高,HIF-1α的表达变化可能影响细胞的存活水平。Objective To study the regulation of hypoxia inducible factor-1α(HIF-1α) expression in hepatoma cells after irradiation and the expression of HIF-1α effect on the radiosensitivity of heptoma cells. Methods HepG2 cells were pretreated by Cobalt chloride (COCl2), a chemical hypoxia agent, to induce and stabilize the expression of HIF-1α, and then exposed to different γ-irradiation doses. Clonogenic assay was used to evaluate HepG2 cell survival fraction (SF) after irradiation under normoxia and chemical hypoxia. Reverse transcriptase polymerase chain reaction (RT-PCR) and immunoblot assay (Western blot) were utilized to detect the changes of intracellular HIF-1α on the level of transcripation and translation, Results Cell survival level was elevated by chemical hypoxia and there was a statistical difference between chemical hypoxic group and normoxic group. The ratios of SF( SFco/SFo2 )on two different conditions were increased with irradiation doses. Meanwhile, the irradiation induced up-regulation of HIF-1α in dose-dependent manner. The expression of HIF-1α was correlated with HepG2 cell survival level to some extent. Conclusions Irradiation could up-regulate the level of HIF-1α expression in HepG2 cells under chemical hypoxic condition. The cells survival level might be influenced by the changes in HIF-1α expression.
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