發(fā)布時(shí)間：2019-03-30 16:12

【摘要】：目的高遷移率族蛋白B1(HMGB1)通過(guò)影響線粒體自噬參與腫瘤細(xì)胞能量代謝,文中旨在探討缺氧環(huán)境下HMGB1對(duì)線粒體生物合成以及細(xì)胞能量代謝的影響。方法 HepG2細(xì)胞設(shè)常氧對(duì)照組(細(xì)胞在含5%CO_2的正常培養(yǎng)箱中培養(yǎng))、缺氧對(duì)照組(細(xì)胞在1%O2+5%CO_2+94%N_2三氣培養(yǎng)箱中培養(yǎng))、HMGB1siRNA缺氧組(細(xì)胞轉(zhuǎn)染HMGB1siRNA后于1%O2+5%CO_2+94%N_2三氣培養(yǎng)箱中培養(yǎng))和siRNA缺氧對(duì)照組(細(xì)胞轉(zhuǎn)染陰性對(duì)照siRNA后于1%O2+5%CO_2+94%N_2三氣培養(yǎng)箱中培養(yǎng))。MTS法檢測(cè)各組細(xì)胞增殖的速度,RT-PCR和Western blot檢測(cè)各組細(xì)胞線粒體生物合成相關(guān)分子表達(dá)變化,透射電鏡觀察細(xì)胞內(nèi)線粒體形態(tài)和數(shù)量,ATP試劑盒檢測(cè)細(xì)胞內(nèi)ATP含量。結(jié)果 HMGB1siRNA缺氧組細(xì)胞在48 h、72 h的吸光度值明顯低于缺氧對(duì)照組和siRNA缺氧對(duì)照組(P0.05)。當(dāng)HMGB1表達(dá)被抑制后,PGC1α、NRF1和TFAM的相對(duì)表達(dá)量明顯低于缺氧對(duì)照組和siRNA缺氧對(duì)照組(P0.05)。Western blot結(jié)果顯示,在缺氧培養(yǎng)24 h后,PGC1α、NRF1和TFAM蛋白的相對(duì)表達(dá)量明顯高于常氧對(duì)照組(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)。當(dāng)HMGB1表達(dá)被抑制后,PGC1α、NRF1和TFAM蛋白的相對(duì)表達(dá)量明顯低于缺氧對(duì)照組和siRNA缺氧對(duì)照組(P0.05)。與缺氧對(duì)照組相比,HMGB1 siRNA缺氧組細(xì)胞內(nèi)ATP含量明顯下降,以缺氧12 h和24 h最為明顯(P0.05)。結(jié)論 HMGB1通過(guò)調(diào)控線粒體生物合成,維持細(xì)胞能量代謝,使細(xì)胞在不利于自身生長(zhǎng)的缺氧環(huán)境下繼續(xù)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 偽, 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 鹵0.210 vs 0.090 鹵0.020, 1.080 鹵0.470 vs 0.581 鹵0.190, 1.585 鹵0.340 vs 0.792 鹵0.350, P0.05). When the expression of HMGB1 was inhibited, the relative expression of PGC1 偽, 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]
【作者單位】： 贛南醫(yī)學(xué)院第一附屬醫(yī)院內(nèi)分泌科;贛南醫(yī)學(xué)院第一附屬醫(yī)院醫(yī)務(wù)科;贛南醫(yī)學(xué)院第一附屬醫(yī)院消化科;
【基金】：國(guó)家自然科學(xué)基金(81660406) 江西省衛(wèi)生計(jì)生委科技計(jì)劃項(xiàng)目(20161096) 贛州市指導(dǎo)性科技計(jì)劃(社會(huì)發(fā)展)項(xiàng)目(GZ2015ZSF072) 贛南醫(yī)學(xué)院校級(jí)重點(diǎn)科研課題(ZD201601)
【分類號(hào)】：R735.7

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