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Mitoferrin-1(MFRN1)是溶质携带蛋白家族的一员,是线粒体内膜上最主要的铁运输蛋白。为了观察MFRN1过表达对293T细胞生长及线粒体功能的影响,采用包装慢病毒和感染293T细胞的方法构建MFRN1稳定过表达的细胞模型(293T-MFRN1),并对其线粒体功能进行评价。构建的稳定细胞株293T-MFRN1中MFRN1转录水平及蛋白表达水平显著增加(P<0.01),MFRN1过表达细胞线粒体内总铁的含量明显增加(P<0.05);电镜显示线粒体结构损伤,线粒体嵴减少、空泡化;线粒体能量代谢受抑制,细胞ATP含量显著下降,腺苷酸池总量(ATP+ADP+AMP)比对照组减少约40%;细胞生长受到抑制,在细胞培养48 h后高表达组细胞数量显著低于对照组;细胞内羟自由基(·OH)含量增加;线粒体DNA发生明显的氧化损伤。以上表明,MFRN1表达量的稳定对细胞功能极为重要,表达量异常增加将导致细胞线粒体功能障碍、细胞增殖抑制。
Mitoferrin-1 (MFRN1) is a member of the solute-carrying protein family and is the most important iron transporter protein in the mitochondrial inner membrane. To observe the effect of MFRN1 overexpression on the growth and mitochondrial function of 293T cells, a 293T-MFRN1 cell line stably overexpressing MFRN1 was constructed by packaging lentivirus and infecting 293T cells. Mitochondrial function was also evaluated. The mRNA and protein expression of MFRN1 in 293T-MFRN1 cells was significantly increased (P <0.01), and the content of total iron in MFRN1 overexpression cells was significantly increased (P <0.05). The results of electron microscopy showed that mitochondrial structure damage, mitochondrial cristae (ATP + ADP + AMP) decreased by about 40% compared with the control group; cell growth was inhibited, and after 48 h of cell culture The number of cells in the high expression group was significantly lower than that in the control group. The content of hydroxyl radical (· OH) in the cells increased, and the oxidative damage of mitochondrial DNA was observed. The above shows that the stability of the expression of MFRN1 is extremely important to the cell function. The abnormal increase of the expression will lead to cell mitochondrial dysfunction and cell proliferation inhibition.