Apoptosis of human breast carcinoma cells in the presence of cis-platin and L-/D-PPMP: IV. Modulation of replication complexes and glycolipid: Glycosyltransferases.
Apoptosis of human breast carcinoma cells in the presence of cis-platin and L-/D-PPMP: IV. Modulation of replication complexes and glycolipid: Glycosyltransferases.
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顺铂和 L-/D-PPMP 存在下人乳腺癌细胞的凋亡：IV。
DOI：
10.1007/s10719-006-7923-5
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发表时间：
2006
影响因子：
3
通讯作者：
Basu,Subhash
中科院分区：
文献类型：
--
作者：
Boyle,PatrickJ;Ma,Rui;Tuteja,Narendra;Banerjee,Sipra;Basu,Subhash
人乳腺癌细胞（SKBR-3、MCF-7和MDA-468）在分别用抗癌药物顺铂和鞘糖脂生物合成抑制剂L-和D-PPMP处理后观察到凋亡。这些药物以剂量依赖的方式启动细胞凋亡，通过表型形态学变化，荧光磷脂酰丝氨酸特异性染料（PSS-380）结合到细胞膜的外小叶上，以及激活半胱天冬酶，−3，−8和−9来测量。观察到在两小时内，很少有凋亡过程开始，但主要的生化变化发生在6小时后。DNA降解在药物处理24小时后开始。然而，很少有人知道在凋亡过程中的“复制复合物”的稳定性。DNA解旋酶是在基因组DNA复制、修复和重组过程中催化基因组DNA解链的马达蛋白。以前，DNA解旋酶-III的特点是作为复制复合物的一个组成部分，从鸡胚脑以及乳腺癌和结肠癌细胞分离。解旋酶活性通过一种新方法（罗马测定法）测定，DNA聚合酶-α活性通过切口ACT-DNA的常规链延伸测定，通过测定从+/− aphidicolin处理的孵育混合物中获得的值。在所有三种乳腺癌细胞系中，观察到一个共同的趋势：DNA聚合酶-α和解旋酶III活性降低。L-PPMP处理的凋亡癌细胞中糖脂唾液酸转移酶SAT-2（CMP-NeuAc; GD 3 α2-8唾液酸转移酶）和SAT-4（CMP-NeuAc：GM 1a α2-3唾液酸转移酶）的活性明显低于顺铂。
Apoptosis of human breast carcinoma cells (SKBR-3, MCF-7, and MDA-468) has been observed after treatment of these cells with anti-cancer drug cis-platin and glycosphingolipid biosynthesis inhibitor L- and D-PPMP, respectively. These drugs initiated apoptosis in a dose-dependent manner as measured by phenotypic morphological changes, by binding of a fluorescent phophatidyl serine-specific dye (PSS-380) onto the outer leaflet of the cell membranes, and by activation of caspases, −3, −8, and −9. It was observed that in two hours very little apoptotic process had started but predominant biochemical changes occurred after 6 h. DNA degradation started after 24 hours of drug treatment. However, very little is known about the stability of the ‘`Replication Complexes’’ during the apoptotic process. DNA helicases are motor proteins that catalyze the melting of genomic DNA during its replication, repair, and recombination processes. Previously, DNA helicase-III was characterized as a component of the replication complexes isolated from embryonic chicken brains as well as breast and colon carcinoma cells. Helicase activities were measured by a novel method (ROME assay), and DNA polymerase-α activities were determined by regular chain extension of the nicked ACT-DNA, by determining values obtained from +/− aphidicolin-treated incubation mixtures. In all three breast carcinoma cell lines, a common trend was observed: a decrease of activities of DNA polymerase-α and Helicase III. A sharp decrease of activities of the glycolipid sialyltransferases: SAT-2 (CMP-NeuAc; GD3 α2-8 sialyltransferase) and SAT-4 (CMP-NeuAc: GM1a α2-3 sialyltransferase) was observed in the apoptotic carcinoma cells treated with L-PPMP compared withcis-platin.