缺氧诱导因子-1和凋亡信号调节激酶1在肝癌细胞SMMC-7721缺氧时化疗耐药形成中的作用  

Study on the function of HIF-1 and ASK1 in the formation of chemotherapy resistance of hepatoma cell line SMMC-7721 during hypoxia

作　　者：金晶[1] 王伟林[1] 姚敏亚[1] 谢海洋[1] 殷胜勇 周琳[1] 郑树森[1] 

机构地区：[1]浙江大学医学院附属第一医院肝胆胰外科卫生部多器官联合移植研究重点实验室,杭州310003

出　　处：《中华实验外科杂志》2008年第2期206-208,共3页Chinese Journal of Experimental Surgery

基　　金：国家重点基础研究发展计划资助项目（2003CB515501）;浙江省自然科学基金资助项目（Y205173）

摘　　要：目的研究不同氧气条件下肝癌细胞HIE-1和ASK1表达水平在化疗过程中的变化。方法噻唑蓝（MTT）比色法检测肝癌细胞生长情况和药物中效浓度（IC50）.逆转录-聚合酶链反应（RT-PCR）和Western blot法检测HIF—1α和ASK1表达水平，比较氧气、化疗药物、HIF-1抑制等因素对他们表达的影响。结果化疗对肝癌细胞常氧时抑制效果较缺氧时更明显，常氧和缺氧IC50存在明显差异。缺氧时两者表达水平均升高，HIF-1α在IC50缺氧组中翻译水平最高，ASK1在IC50常氧组中翻译表达最多。结论HIF-1α可能在缺氧时通过对ASK1蛋白水平的抑制实现肝癌细胞化疗耐药形成。Objective To investigate the difference of the oxaliplatin therapy effect to human hepatoma cells SMMC-7721 between the condition of hypoxia and normoxia. And to explore the chemotherapy resistance mechanism through the observation of transcription and translation level of HIF-1α and ASK1 on an apoptosis hypothesis. Methods MTT assay was used to measure human hepatoma cells SMMC-7721 proliferation after 24 h growth under normoxia or hypoxia with different oxaliplatin ultimate concentrations （16 μmoL/L,32 μmoL/L.64 μmoL/L. 128 μmoL/L and 256 μmoL/L）. The transcription and translation levels of HIF-1α and ASK1 were measured using RT-PCR and Western blotting. Among the control group （C） ,oxaliplatin-normoxia group （ON） ,oxaliplatin-hypoxia group （OH） and oxaliplatin-hypoxia-YC group （OHY） ,the genes translation level was compared. Results The suppression effect was observed at different drug concentrations in MTT tests, which got much more significant as the drug concentration elevated and was more significant under normoxia. The 50% cell-inhibition concentrations for normoxia and hypoxia were 85.9 μmoL/L and 226.1 μmoL/L （ P 〈 0.05 ）. The transcription levels of HIF- 1α and ASK1 elevated as the hypoxia lasted. The translation level of HIF-1α acted as its transcription level,but ASK1 got to the peak at 12h. The highest translation level of HIF-1α and ASK1 was observed in the OH and ON group, respectively. Conclusions Hypoxia condition helps to form the chemotherapy resistanee. HIF-1 participates the apoptosis process indirectly, which attributes to decrease the chemotherapy effect under hypoxia by suppressing the ASK1 at the level of translation.

关 键 词：缺氧诱导因子1 凋亡信号调节激酶1 缺氧 凋亡 化疗耐药 

分 类 号：R735.7[医药卫生—肿瘤] R364.4[医药卫生—临床医学]