Cyclin D1 cooperates with p21 to regulate TGFβ-mediated breast cancer cell migration and tumor local invasion.
Cyclin D1 cooperates with p21 to regulate TGFβ-mediated breast cancer cell migration and tumor local invasion.
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DOI：
10.1186/bcr3441
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发表时间：
2013-06-20
期刊：
影响因子：
--
通讯作者：
Lebrun JJ
中科院分区：
文献类型：
--
作者：
Dai M;Al-Odaini AA;Fils-Aimé N;Villatoro MA;Guo J;Arakelian A;Rabbani SA;Ali S;Lebrun JJ
人类癌症中经常发现细胞周期机制失调。细胞周期调节剂功能和表达的调节不仅导致增殖优势，而且导致肿瘤进展和癌症的侵袭性。特别是，细胞周期蛋白 D1 和 p21 在人类癌症中经常过度表达，与肿瘤分级高、预后不良和转移增加相关。这促使我们研究转化生长因子β (TGFβ) 下游的细胞周期蛋白 D1/p21 信号轴在乳腺癌进展中的作用。通过定量实时 PCR 和蛋白质印迹法评估三阴性乳腺癌细胞系中细胞周期蛋白 mRNA 和蛋白的表达。细胞周期蛋白 D1 和 p21 之间的共定位和相互作用分别通过免疫细胞化学和免疫共沉淀进行。通过伤口愈合和定量延时成像测定来评估细胞迁移。此外，通过用F-肌动蛋白标记物鬼笔环肽和间充质中间丝波形蛋白染色来检查细胞周期蛋白D1对细胞结构和肌动蛋白组织的影响。最后，使用乳腺脂肪垫异种移植小鼠模型来评估乳腺肿瘤生长和局部侵袭。我们发现 TGFβ 可以特异性上调三阴性乳腺癌细胞中细胞周期蛋白 D1 的表达。 TGFβ 介导的细胞迁移也需要细胞周期蛋白 D1 的诱导。细胞周期蛋白 D1 表达的抑制不仅导致圆形和上皮样表型，而且还阻止 TGFβ 诱导的波形蛋白和 F-肌动蛋白在细胞边缘的共定位以及侵袭伪足的形成。此外，TGFβ促进细胞周期蛋白D1和p21之间的核共定位和物理相互作用。细胞周期蛋白 D1 和 p21 蛋白的共表达是肿瘤发展的初始步骤所必需的，因为这两种分子的双重敲低可阻止异种移植小鼠模型中原发性肿瘤的形成。此外，体内研究表明，在缺乏细胞周期蛋白 D1 和 p21 的情况下，侵袭性肿瘤的局部晚期特征，包括骨骼肌、乳腺脂肪垫和淋巴管侵犯以及溃疡皮肤，都会减弱。因此，我们的研究结果强调了细胞周期蛋白 D1/p21 信号轴作为 TGFβ 介导的肿瘤生长起始和局部肿瘤细胞侵袭的关键调节因子，无论是在体外还是在体内。
Deregulation of the cell cycle machinery is often found in human cancers. Modulations in the cell cycle regulator function and expression result not only in proliferative advantages, but also lead to tumor progression and invasiveness of the cancer. In particular, cyclin D1 and p21 are often over-expressed in human cancers, correlating with high tumor grade, poor prognosis and increased metastasis. This prompted us to investigate the role of the cyclin D1/p21 signaling axis downstream of transforming growth factor beta (TGFβ) in breast cancer progression. Cyclins mRNA and protein expressions were assessed by quantitative real-time PCR and Western blot in triple negative breast cancer cell lines. Co-localization and interaction between cyclin D1 and p21 were performed by immunocytochemistry and co-immunoprecipitation, respectively. Cell migration was assessed by wound healing and quantitative time-lapse imaging assays. In addition, the effects of cyclin D1 on cellular structure and actin organization were examined by staining with F-actin marker phalloidin and mesenchymal intermediate filament vimentin. Finally, a mammary fat pad xenograft mouse model was used to assess mammary tumor growth and local invasion. We found TGFβ to specifically up-regulate the expression of cyclin D1 in triple negative breast cancer cells. Induction of cyclin D1 is also required for TGFβ-mediated cell migration. Suppression of cyclin D1 expression not only resulted in a rounded and epithelial-like phenotype, but also prevented TGFβ-induced vimentin and F-actin co-localization at the cell edge as well as invadopodia formation. Furthermore, TGFβ promoted the nuclear co-localization and physical interaction between cyclin D1 and p21. The co-expression of cyclin D1 and p21 proteins are required for the initial steps of tumor development, as double knockdown of these two molecules prevented primary tumor formation in a Xenograft mouse model. Moreover, the in vivo studies indicated that locally advanced features of the invasive tumors, including skeletal muscle, mammary fat pad and lymphovascular invasion, as well as ulcerated skin, were attenuated in the absence of cyclin D1 and p21. Thus, our findings highlight the cyclin D1/p21 signaling axis as a critical regulator of TGFβ-mediated tumor growth initiation and local tumor cell invasion, both in vitro and in vivo.
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DOI：
10.1158/1078-0432.ccr-08-3293
发表时间：
2009-06-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Agarwal R;Gonzalez-Angulo AM;Myhre S;Carey M;Lee JS;Overgaard J;Alsner J;Stemke-Hale K;Lluch A;Neve RM;Kuo WL;Sorlie T;Sahin A;Valero V;Keyomarsi K;Gray JW;Borresen-Dale AL;Mills GB;Hennessy BT
通讯作者：
Hennessy BT
影响因子：
64.5
作者：
ELDEIRY, WS;TOKINO, T;VOGELSTEIN, B
通讯作者：
VOGELSTEIN, B
影响因子：
64.8
作者：
Jirawatnotai S;Hu Y;Michowski W;Elias JE;Becks L;Bienvenu F;Zagozdzon A;Goswami T;Wang YE;Clark AB;Kunkel TA;van Harn T;Xia B;Correll M;Quackenbush J;Livingston DM;Gygi SP;Sicinski P
通讯作者：
Sicinski P
影响因子：
16
作者：
Chacko, BM;Qin, BY;Lin, K
通讯作者：
Lin, K
DOI：
10.1038/84995
发表时间：
2001-03-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Chacko, BM;Qin, B;Lin, K
通讯作者：
Lin, K