Over-expression of pemt2 into Rat Hepatoma Cells Contributes to the Mitochondrial Apoptotic Pathway
Over-expression of pemt2 into Rat Hepatoma Cells Contributes to the Mitochondrial Apoptotic Pathway
复制标题
pemt2 在大鼠肝癌细胞中的过表达有助于线粒体凋亡途径
DOI：
10.1002/iub.222
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发表时间：
2009-08-01
期刊：
影响因子：
4.6
通讯作者：
Ma, Keli
中科院分区：
文献类型：
--
作者：
Li, Yali;Zou, Wei;Ma, Keli
我们之前建立了一个磷脂酰乙醇胺N-甲基转移酶2（pemt 2）稳定转染CBRH-7919肝癌细胞系，并表明pemt 2过表达抑制细胞增殖并诱导凋亡。本研究旨在进一步阐明导致这些细胞凋亡的细胞机制。采用脂质提取、高效薄层色谱、高效气相色谱和[H-3]-乙醇胺示踪等方法分析了pemt 2过表达细胞和对照细胞中磷脂酰胆碱（PC）的脂肪酸组成以及PEMT 2途径合成PC的位置。Western blot检测pemt 2过表达对线粒体膜流动性、细胞色素C释放和caspase活性的影响。PEMT 2新合成的PC含有较多的油酸酰基（P <0. 01），主要位于线粒体; pemt 2过表达增加线粒体膜流动性，细胞色素C从线粒体释放到胞浆，进而激活线粒体凋亡途径中的关键分子caspase-9和caspase-3。我们证明，在大鼠肝癌细胞中，PEMT 2诱导的凋亡通过线粒体进行。(C)2009 IUBMB IUBMB Life，61（8）：846-852，2009
We previously established a line of phosphatidylethanolamine N-methyltransferase 2 (pemt2) -stably transfected CBRH-7919 hepatoma cells, and showed that pemt2 over-expression inhibited cell proliferation and induced apoptosis. This study was aimed to further elucidate the cellular mechanisms leading to this apoptosis in these cells. Fatty acid compositions of phosphatidylcholine (PC) in pemt2 over-expressed cells and control cells, and the location of PC synthesized by PEMT2 pathway were analyzed with lipid extraction, high-performance thin layer chromatography, high-performance gas chromatography (HPGC), and [H-3]-ethanolamine tracing. The effects of pemt2 overexpression on the mitochondrial membrane fluidity, the release of cytochrome C from mitochondria, and the activity of caspases were determined by Western blot. Newly synthesized PC by PEMT2 contained more acyl groups of oleic acid (P < 0.01) and was mainly located in mitochondria; pemt2 over-expression increased the mitochondrial membrane fluidity and the release of cytochrome C from the mitochondria into the cytoplasma, which in turn activated caspase-9 and caspase-3, the key molecules in the mitochondrial apoptotic pathway. We demonstrated that, in rat hepatoma cells, PEMT2-induced apoptosis proceeds through mitochondria. (C) 2009 IUBMB IUBMB Life, 61(8): 846-852, 2009