缺氧环境下高迁移率族蛋白B1对线粒体生物合成的影响
发布时间：2019-03-30 16:12
【摘要】：目的高迁移率族蛋白B1(HMGB1)通过影响线粒体自噬参与肿瘤细胞能量代谢,文中旨在探讨缺氧环境下HMGB1对线粒体生物合成以及细胞能量代谢的影响。方法 HepG2细胞设常氧对照组(细胞在含5%CO_2的正常培养箱中培养)、缺氧对照组(细胞在1%O2+5%CO_2+94%N_2三气培养箱中培养)、HMGB1siRNA缺氧组(细胞转染HMGB1siRNA后于1%O2+5%CO_2+94%N_2三气培养箱中培养)和siRNA缺氧对照组(细胞转染阴性对照siRNA后于1%O2+5%CO_2+94%N_2三气培养箱中培养)。MTS法检测各组细胞增殖的速度,RT-PCR和Western blot检测各组细胞线粒体生物合成相关分子表达变化,透射电镜观察细胞内线粒体形态和数量,ATP试剂盒检测细胞内ATP含量。结果 HMGB1siRNA缺氧组细胞在48 h、72 h的吸光度值明显低于缺氧对照组和siRNA缺氧对照组(P0.05)。当HMGB1表达被抑制后,PGC1α、NRF1和TFAM的相对表达量明显低于缺氧对照组和siRNA缺氧对照组(P0.05)。Western blot结果显示,在缺氧培养24 h后,PGC1α、NRF1和TFAM蛋白的相对表达量明显高于常氧对照组(0.494±0.210 vs 0.090±0.020,1.080±0.470 vs 0.581±0.190,1.585±0.340 vs 0.792±0.350,P0.05)。当HMGB1表达被抑制后,PGC1α、NRF1和TFAM蛋白的相对表达量明显低于缺氧对照组和siRNA缺氧对照组(P0.05)。与缺氧对照组相比,HMGB1 siRNA缺氧组细胞内ATP含量明显下降,以缺氧12 h和24 h最为明显(P0.05)。结论 HMGB1通过调控线粒体生物合成,维持细胞能量代谢,使细胞在不利于自身生长的缺氧环境下继续增殖。
[Abstract]:Aim High mobility group B1 (HMGB1) is involved in energy metabolism of tumor cells by affecting mitochondrial autophagy. The aim of this study was to investigate the effects of HMGB1 on mitochondrial biosynthesis and cell energy metabolism in hypoxic environment. Methods HepG2 cells were divided into normoxic control group (cultured in normal culture chamber containing 5%CO_2) and hypoxic control group (cultured in 1%O2 5%CO_2 94% N2 three-gas incubator). HMGB1siRNA hypoxia group (cells transfected with HMGB1siRNA were cultured in 1%O2 5%CO_2 94% N _ 2 three-gas incubator) and siRNA hypoxia control group (cell negative control siRNA was cultured in 1%O2 5%CO_2 94% N _ 2 three-gas medium). The rate of cell proliferation in each group was measured by). MTS method. RT-PCR and Western blot were used to detect the expression of mitochondrial biosynthetic molecules, transmission electron microscopy (TEM) was used to observe the morphology and quantity of mitochondria, and ATP kit was used to detect the content of ATP in cells. Results the absorbance of cells in HMGB1siRNA hypoxia group at 48 h and 72 h was significantly lower than that in hypoxia control group and siRNA hypoxia control group (P0.05). When the expression of HMGB1 was inhibited, the relative expression of PGC1 伪, NRF1 and TFAM was significantly lower than that of hypoxia control group and siRNA hypoxia control group (P0.05). Western blot). The relative expression of NRF1 and TFAM protein was significantly higher than that of normoxic control group (0.494 卤0.210 vs 0.090 卤0.020, 1.080 卤0.470 vs 0.581 卤0.190, 1.585 卤0.340 vs 0.792 卤0.350, P0.05). When the expression of HMGB1 was inhibited, the relative expression of PGC1 伪, NRF1 and TFAM protein was significantly lower than that of hypoxia control group and siRNA hypoxia control group (P0.05). Compared with the hypoxic control group, the intracellular ATP content in HMGB1 siRNA hypoxia group was significantly decreased, especially at 12 h and 24 h after hypoxia (P0.05). Conclusion HMGB1 can maintain cell energy metabolism by regulating mitochondrial biosynthesis, so that cells continue to proliferate in anoxic environment which is not conducive to their own growth. [WT5 "HZ] conclusion [WT5" BZ]
【作者单位】： 赣南医学院第一附属医院内分泌科;赣南医学院第一附属医院医务科;赣南医学院第一附属医院消化科;
【基金】：国家自然科学基金(81660406) 江西省卫生计生委科技计划项目(20161096) 赣州市指导性科技计划(社会发展)项目(GZ2015ZSF072) 赣南医学院校级重点科研课题(ZD201601)
【分类号】：R735.7
[Abstract]:Aim High mobility group B1 (HMGB1) is involved in energy metabolism of tumor cells by affecting mitochondrial autophagy. The aim of this study was to investigate the effects of HMGB1 on mitochondrial biosynthesis and cell energy metabolism in hypoxic environment. Methods HepG2 cells were divided into normoxic control group (cultured in normal culture chamber containing 5%CO_2) and hypoxic control group (cultured in 1%O2 5%CO_2 94% N2 three-gas incubator). HMGB1siRNA hypoxia group (cells transfected with HMGB1siRNA were cultured in 1%O2 5%CO_2 94% N _ 2 three-gas incubator) and siRNA hypoxia control group (cell negative control siRNA was cultured in 1%O2 5%CO_2 94% N _ 2 three-gas medium). The rate of cell proliferation in each group was measured by). MTS method. RT-PCR and Western blot were used to detect the expression of mitochondrial biosynthetic molecules, transmission electron microscopy (TEM) was used to observe the morphology and quantity of mitochondria, and ATP kit was used to detect the content of ATP in cells. Results the absorbance of cells in HMGB1siRNA hypoxia group at 48 h and 72 h was significantly lower than that in hypoxia control group and siRNA hypoxia control group (P0.05). When the expression of HMGB1 was inhibited, the relative expression of PGC1 伪, NRF1 and TFAM was significantly lower than that of hypoxia control group and siRNA hypoxia control group (P0.05). Western blot). The relative expression of NRF1 and TFAM protein was significantly higher than that of normoxic control group (0.494 卤0.210 vs 0.090 卤0.020, 1.080 卤0.470 vs 0.581 卤0.190, 1.585 卤0.340 vs 0.792 卤0.350, P0.05). When the expression of HMGB1 was inhibited, the relative expression of PGC1 伪, NRF1 and TFAM protein was significantly lower than that of hypoxia control group and siRNA hypoxia control group (P0.05). Compared with the hypoxic control group, the intracellular ATP content in HMGB1 siRNA hypoxia group was significantly decreased, especially at 12 h and 24 h after hypoxia (P0.05). Conclusion HMGB1 can maintain cell energy metabolism by regulating mitochondrial biosynthesis, so that cells continue to proliferate in anoxic environment which is not conducive to their own growth. [WT5 "HZ] conclusion [WT5" BZ]
【作者单位】： 赣南医学院第一附属医院内分泌科;赣南医学院第一附属医院医务科;赣南医学院第一附属医院消化科;
【基金】：国家自然科学基金(81660406) 江西省卫生计生委科技计划项目(20161096) 赣州市指导性科技计划(社会发展)项目(GZ2015ZSF072) 赣南医学院校级重点科研课题(ZD201601)
【分类号】：R735.7
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