Reduced expression of β-catenin inhibitor Chibby in colon carcinoma cell lines
Reduced expression of β-catenin inhibitor Chibby in colon carcinoma cell lines
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DOI：
10.3748/wjg.v12.i10.1529
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发表时间：
2006-03-14
影响因子：
4.3
通讯作者：
Bosserhoff, Anja-Katrin
中科院分区：
文献类型：
--
作者：
Schuierer, Marion M.;Graf, Elisabeth;Bosserhoff, Anja-Katrin
目的：目的：分析Chibby在结肠癌细胞系和结直肠癌（CRC）中的表达及其功能。通过测序分析Chibby基因的突变情况。为了测试Chibby是否对结肠癌细胞中的β-连环蛋白信号传导表现出影响，我们用Chibby表达质粒转染SW 480细胞，随后分析β-连环蛋白的活性并测试细胞表型的改变。此外，我们还采用定量RT-PCR和杂交基因芯片技术检测了结直肠癌组织和癌旁正常组织中Chibby mRNA的表达水平。与正常结肠上皮细胞相比，Chibby mRNA表达在结肠癌细胞系中强烈下调，并且在任何检查的结肠癌细胞系中均未发现突变。我们可以证明，当在SW 480细胞中过表达时，Chibby在TOPflash测定中抑制β-连环蛋白活性。与对照细胞相比，用Chibby转染的SW 480细胞进行的增殖和侵袭测定没有显示出显著差异。与这些体外数据相反，CRC肿瘤样品中Chibby mRNA水平的定量RT-PCR分析与邻近非癌组织中的样品没有显示出显著差异。与这些研究结果相一致，基因芯片分析肿瘤组织样本和相应的正常组织没有显示改变Chibby expression.CONCLUSION：改变Chibby表达可能会在体外观察到在不同的结肠癌细胞系。然而，这一发现无法在体外CRC肿瘤中得到证实，表明Chibby不太可能促进CRC肿瘤的发展或进展。由于Chibby是β-连环蛋白信号传导的重要抑制剂，我们的数据暗示，结肠癌细胞系用于分析结直肠癌中Wnt/β-连环蛋白通路的体外研究的可用性需要广泛的验证。(C)2006年，WJG出版社。All rights reserved.
AIM: To analyse the Chibby expression and its function in colon carcinoma cell lines and colorectal carcinoma (CRC).METHODS: Chibby expression levels were investigated by quantitative RT-PCR in a panel of seven different colon carcinoma cell lines. By sequencing, we analysed mutational status of Chibby. To test whether Chibby exhibited effects on beta-catenin signalling in colon carcinoma cells, we transfected SW480 cells with Chibby expression plasmid and, subsequently, analysed activity of beta-catenin and tested for alterations in cellular phenotype. In addition, we examined Chibby mRNA levels in samples of colorectal carcinomas and adjacent normal tissues by using quantitative RT-PCR and hybridised gene chips with samples from CRC and normal tissues.RESULTS: Chibby mRNA expression was strongly down-regulated in colon carcinoma cell lines in comparison to normal colon epithelial cells and no mutation in any of the examined colon carcinoma cell lines was found. Further, we could show that Chibby inhibited beta-catenin activity in TOPflash assays when over-expressed in SW480 cells. Proliferation and invasion assays with Chibby transfected SW480 cells did not reveal profound differences compared to control cells. In contrast to these in vitro data, quantitative RT-PCR analyses of Chibby mRNA levels in CRC tumor samples did not show significant differences to specimens in adjacent non-cancerous tissue. Consistent with these findings, gene chips analysing tissue samples of tumors and corresponding normal tissue did not show altered Chibby expression.CONCLUSION: Altered Chibby expression might be observed in vitro in different colon carcinoma cell lines. However, this finding could not be confirmed in vitro in CRC tumors, indicating that Chibby is not likely to promote CRC tumor development or progression. As Chibby is an important inhibitor of beta-catenin signalling, our data implicate that the usability of colon carcinoma cell lines for in vitro studies analysing the Wnt/beta-catenin pathway in colorectal carcinoma needs extensive verification. (C) 2006 The WJG Press. All rights reserved.