Plasma proteomics of pancreatic cancer patients by multi-dimensional liquid chromatography and two-dimensional difference gel electrophoresis (2D-DIGE): up-regulation of leucine-rich alpha-2-glycoprotein in pancreatic cancer.
Plasma proteomics of pancreatic cancer patients by multi-dimensional liquid chromatography and two-dimensional difference gel electrophoresis (2D-DIGE): up-regulation of leucine-rich alpha-2-glycoprotein in pancreatic cancer.
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DOI：
10.1016/j.jchromb.2007.01.029
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发表时间：
2007-06-01
期刊：
影响因子：
--
通讯作者：
Hirohashi S
中科院分区：
文献类型：
--
作者：
Kakisaka T;Kondo T;Okano T;Fujii K;Honda K;Endo M;Tsuchida A;Aoki T;Itoi T;Moriyasu F;Yamada T;Kato H;Nishimura T;Todo S;Hirohashi S
我们研究了胰腺癌患者血浆蛋白的异常表达。高丰度的血浆蛋白（白蛋白，转铁蛋白，触珠蛋白，α-1-抗胰蛋白酶，IgG和伊加）被耗尽通过使用免疫亲和柱，和低丰度的分离成五个馏分通过阴离子交换色谱。将分级分离的血浆蛋白用高灵敏度荧光染料进行2D-DIGE。通过2D-DIGE获得的定量蛋白质表达谱在两种血浆蛋白质混合物之间进行比较：一种来自五名非癌症健康供体，另一种来自五名胰腺癌患者。在1200个蛋白质点中，我们发现33个蛋白质点在两种混合物之间表达不同;其中27个蛋白质点在癌症中上调，6个蛋白质点下调。质谱和数据库搜索允许鉴定对应于凝胶点的蛋白质。观察到富含亮氨酸的α-2-糖蛋白（LRG）的上调，该蛋白以前未涉及胰腺癌。用抗LRG抗体的蛋白质印迹法验证了来自健康对照、慢性胰腺炎患者和胰腺癌患者的独立系列血浆样品中LRG的上调。我们的研究结果证明了多维液相色谱与2D-DIGE相结合在血浆蛋白质组学中的应用，并表明LRG血浆水平测量的临床实用性。
We investigated the aberrant expression of plasma proteins in patients with pancreatic cancer. High-abundance plasma proteins (albumin, transferrin, haptoglobin, alpha-1-antitrypsin, IgG and IgA) were depleted by use of an immuno-affinity column, and low-abundance ones were separated into five fractions by anion-exchange chromatography. The fractionated plasma proteins were subjected to 2D-DIGE with highly sensitive fluorescent dyes. The quantitative protein expression profiles obtained by 2D-DIGE were compared between two plasma protein mixtures: one from five non-cancer bearing healthy donors and the other from five patients with pancreatic cancer. Among 1200 protein spots, we found that 33 protein spots were differently expressed between the two mixtures; 27 of these were up-regulated and six were down-regulated in cancer. Mass spectrometry and database searching allowed the identification of the proteins corresponding to the gel spots. Up-regulation of leucine-rich alpha-2-glycoprotein (LRG), which has not previously been implicated in pancreatic cancer, was observed. Western blotting with an anti-LRG antibody validated the up-regulation of LRG in an independent series of plasma samples from healthy controls, patients with chronic pancreatitis, and patients with pancreatic cancer. Our results demonstrate the application of a combination of multi-dimensional liquid chromatography with 2D-DIGE for plasma proteomics and suggest the clinical utility of LRG plasma level measurement.
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影响因子：
3.4
作者：
Fujii, K;Kondo, T;Hirohashi, S
通讯作者：
Hirohashi, S
影响因子：
7
作者：
Gronborg, M;Kristiansen, TZ;Pandey, A
通讯作者：
Pandey, A
影响因子：
2.9
作者：
Bini, L;Magi, B;Pallini, V
通讯作者：
Pallini, V
影响因子：
3.4
作者：
Moritz, RL;Clippingdale, AB;Simpson, RJ
通讯作者：
Simpson, RJ
影响因子：
3.4
作者：
Pieper, R;Su, Q;Steiner, S
通讯作者：
Steiner, S