APC/β-catenin-rich complexes at membrane protrusions regulate mammary tumor cell migration and mesenchymal morphology.
APC/β-catenin-rich complexes at membrane protrusions regulate mammary tumor cell migration and mesenchymal morphology.
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DOI：
10.1186/1471-2407-13-12
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发表时间：
2013-01-09
期刊：
影响因子：
3.8
通讯作者：
Goss KH
中科院分区：
文献类型：
--
作者：
Odenwald MA;Prosperi JR;Goss KH
APC肿瘤抑制因子在许多肿瘤类型中突变或下调，并且显著地定位于迁移细胞中突起尖端处的点状簇，例如在星形胶质细胞中，其中其与定向细胞运动有关。例如，尽管APC丢失被认为是结直肠癌的起始事件，但尚不清楚APC在肿瘤细胞运动中起什么作用，以及APC丢失是否可能是肿瘤进展的重要促进因素。在多种细胞系中分析APC和β-连环蛋白的定位，包括用蛋白酶体抑制剂或TGFβ处理以诱导上皮向间充质转化（EMT）的非转化上皮系，以及几种乳腺癌系，通过免疫荧光。使用APC特异性短发夹（sh）RNA的慢病毒介导的递送在4 T07乳腺肿瘤细胞中敲低APC表达，并使用定量（q）逆转录酶（RT）-PCR和蛋白质印迹法评估。通过进行伤口填充试验分析肿瘤细胞运动性，并通过免疫荧光（IF）和相差显微镜观察形态学。此外，使用BrdU掺入测量增殖，并进行TCF报告基因测定以确定β-连环蛋白/TCF介导的转录活性。APC/β-catenin丰富的复合物被观察到在用蛋白酶体抑制剂处理的迁移性上皮细胞的突起末端处或当EMT已经被诱导时，以及在具有间充质、纺锤形形态的肿瘤细胞中。APC水平降低的4 T07肿瘤细胞的运动性显著降低，并且具有更圆的形态;然而，它们在增殖或β-连环蛋白/TCF转录活性方面没有显著差异。此外，我们发现APC/β-catenin丰富的复合物在突起末端依赖于一个完整的微管细胞骨架。这些发现表明，具有含APC/β-连环蛋白斑点的膜突起控制乳腺肿瘤细胞的迁移潜力和间充质形态，并表明肿瘤进展后期阶段的APC丢失可能会影响肿瘤细胞的传播或定殖。
The APC tumor suppressor is mutated or downregulated in many tumor types, and is prominently localized to punctate clusters at protrusion tips in migratory cells, such as in astrocytes where it has been implicated in directed cell motility. Although APC loss is considered an initiating event in colorectal cancer, for example, it is less clear what role APC plays in tumor cell motility and whether loss of APC might be an important promoter of tumor progression in addition to initiation. The localization of APC and β-catenin was analyzed in multiple cell lines, including non-transformed epithelial lines treated with a proteasome inhibitor or TGFβ to induce an epithelial-to-mesenchymal transition (EMT), as well as several breast cancer lines, by immunofluorescence. APC expression was knocked down in 4T07 mammary tumor cells using lentiviral-mediated delivery of APC-specific short-hairpin (sh) RNAs, and assessed using quantitative (q) reverse-transcriptase (RT)-PCR and western blotting. Tumor cell motility was analyzed by performing wound-filling assays, and morphology via immunofluorescence (IF) and phase-contrast microscopy. Additionally, proliferation was measured using BrdU incorporation, and TCF reporter assays were performed to determine β-catenin/TCF-mediated transcriptional activity. APC/β-catenin-rich complexes were observed at protrusion ends of migratory epithelial cells treated with a proteasome inhibitor or when EMT has been induced and in tumor cells with a mesenchymal, spindle-like morphology. 4T07 tumor cells with reduced APC levels were significantly less motile and had a more rounded morphology; yet, they did not differ significantly in proliferation or β-catenin/TCF transcriptional activity. Furthermore, we found that APC/β-catenin-rich complexes at protrusion ends were dependent upon an intact microtubule cytoskeleton. These findings indicate that membrane protrusions with APC/β-catenin-containing puncta control the migratory potential and mesenchymal morphology of mammary tumor cells and suggest that APC loss during later stages of tumor progression might impact tumor cell dissemination or colonization.
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影响因子：
64.5
作者：
GRODEN, J;THLIVERIS, A;WHITE, R
通讯作者：
WHITE, R
影响因子：
8
作者：
Morrison, EE;Wardleworth, BN;Meredith, DM
通讯作者：
Meredith, DM
影响因子：
29.4
作者：
Heinen, CD;Goss, KH;Groden, J
通讯作者：
Groden, J
影响因子：
3.3
作者：
Kita, K;Wittmann, T;Waterman-Storer, CM
通讯作者：
Waterman-Storer, CM
DOI：
10.1083/jcb.148.3.505
发表时间：
2000-02-07
期刊：
The Journal of cell biology
影响因子：
--
作者：
Mimori-Kiyosue Y;Shiina N;Tsukita S
通讯作者：
Tsukita S