Quantitative cell signalling analysis reveals down-regulation of MAPK pathway activation in colorectal cancer
Quantitative cell signalling analysis reveals down-regulation of MAPK pathway activation in colorectal cancer
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DOI：
10.1002/path.2561
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发表时间：
2009-08-01
影响因子：
7.3
通讯作者：
Petricoin, Emanuel F., III
中科院分区：
文献类型：
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作者：
Gulmann, Christian;Sheehan, Katherine M.;Petricoin, Emanuel F., III
丝裂原活化蛋白激酶（Mitogen-activated protein kinases，MAPK）被认为在结肠癌的发生中起重要作用，激酶抑制剂治疗已被认为是治疗结肠癌的潜在工具。使用磷酸化特异性抗体的反相微阵列测定可以直接测量磷酸化蛋白质亚型的水平。在当前的研究中，对35例未经治疗的结直肠癌结肠切除术的样本进行激光捕获显微切割，以从癌症以及正常（即未受累）粘膜中分离上皮和基质。将从这四种组织类型产生的裂解物点样到反相蛋白微阵列上，并用一组ERK、p-ERK、p38、p-p38、p-JNK、MEK和p-MEK的抗体进行探测。尽管总蛋白水平在癌症中没有变化，或略有升高（p38，p = 0.0025），但与未受累的粘膜相比，癌症中活化的同种型，包括p-ERK、p-p38和p-JNK，降低了2 - 4倍（除了上皮中的p-JNK外，所有病例中p < 0.0023，其中降低不显著）。这得到了蛋白质印迹法的支持。Dukes的B期和C期癌症显示出比Dukes的A期癌症更低的p-ERK和p-p38表达，尽管这在统计学上不显著。它的结论是，MAPK活性可能会下调，在结直肠癌和抑制治疗在这个系统中的进一步探索，应仔细评估，如果这一发现证实了在更大的系列。版权所有（C）2009大不列颠和爱尔兰病理学会。出版社：John Wiley & Sons，Ltd
Mitogen-activated protein kinases (MAPK) are considered to play significant roles in colonic carcinogenesis and kinase inhibitor therapy has been proposed as a potential tool in the treatment of this disease. Reverse-phase microarray assays using phospho-specific antibodies can directly measure levels of phosphorylated protein isoforms. In the current study, samples from 35 cases of untreated colorectal cancer colectomies were laser capture-microdissected to isolate epithelium and stroma from cancer as well as normal (i.e. uninvolved) mucosa. Lysates generated from these four tissue types were spotted onto reverse-phase protein microarrays and probed with a panel of antibodies to ERK, p-ERK, p38, p-p38, p-JNK, MEK and p-MEK. Whereas total protein levels were unchanged, or slightly elevated (p38, p = 0.0025) in cancers, activated isoforms, including p-ERK, p-p38 and p-JNK, were decreased two- to four-fold in cancers compared with uninvolved mucosa (p < 0.0023 in all cases except for p-JNK in epithelium, where decrement was non-significant). This was backed up by western blotting. Dukes' stage B and C cancers displayed lower p-ERK and p-p38 expression than Dukes' stage A cancers, although this was not statistically significant. It is concluded that MAPK activity may be down-regulated in colorectal cancer and that further exploration of inhibitory therapy in this system should be carefully evaluated if this finding is confirmed in larger series. Copyright (C) 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.