HIF-3α在线粒体的表达及意义

李露; 周彦妮; 李胜富; 龙丹; 李鸿生; 冯莉; 李幼平

HIF-3α在线粒体的表达及意义

Expression and Significance of HIF-3α in Mitochondria

摘要

摘要:
目的探究低氧诱导因子-3α(HIF-3α)定位至线粒体的现象及其生理、病理意义。
方法 ① 低氧(1%O₂)或脯氨酸羟化酶抑制剂——二甲基乙二酰甘氨酸(DMOG)、氯化钴(CoCl₂)模拟低氧处理人宫颈癌细胞系HeLa和人肾癌细胞系ACHN后，蛋白质免疫印迹(Western blot)和免疫荧光(IF)检测HIF-3α在细胞中的分布及其在线粒体中的表达；②通过酶敏感实验观察HIF-3α的线粒体亚结构定位；③采用Western blot检测HIF-3α在正常小鼠不同器官组织、人肝癌组织线粒体中的表达情况。
结果 ① 在检测的2种细胞系(HeLa和ACHN细胞)中，HIF-3α在常氧及缺氧条件下均可定位至线粒体，且缺氧组HIF-3α在线粒体中的表达高于常氧组；②HIF-3α的消化模式与线粒体外膜蛋白一致；③HIF-3α在小鼠多种器官组织中均可定位线粒体，HIF-3α在人肝癌组织线粒体中的表达高于正常及癌旁组织。结论HIF-3α在常氧及缺氧条件下均可定位至线粒体外膜，低氧可促进其线粒体定位；该现象可能与肝癌的发生发展相关。

Abstract:
Objective To investigate the mitochondrial translocation of hypoxia inducible factor-3α (HIF-3α) under normoxia and hypoxia and its physiological and pathological meanings.
Methods ① After hypoxic (1%O₂) or DMOG, CoCl₂ treatments mimicking the hypoxic treatment, Western blot and immunofluorescence were used to examine the HIF-3α expression in mitochondria of HeLa and ACHN cells, respectively. ②The protease sensitivity experiment was used to explore the sub-organelle localization of HIF-3α in mitochondria. ③Western blot was used to examine mitochondrial HIF-3α in the normal mouse tissues and human liver carcinoma tissues.
Results ① In HeLa and ACHN cells, HIF-3α translocated to mitochondria under normoxia and hypoxia, and its mitochondrial expression was higher under hypoxia; ②The protease sensitivity of HIF-3α was similar to proteins locating in the mitochondrial outer membrane; ③Mitochondrial HIF-3α expressed in multiple normal mouse tissues; The expression of mitochondrial HIF-3α was higher in human liver carcinoma tissues than the normal and adjacent tissues.
Conclusion HIF-3α translocated to mitochondrial outer membrane under both normoxia and hypoxia, and hypoxia could up-regulated HIF-3α mitochondrial translocation. Meanwhile, the phenomenon may be involved in the process of liver carcinoma.

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