TNFAIP3 is required for FGFR1 activation-promoted proliferation and tumorigenesis of premalignant DCIS.COM human mammary epithelial cells.
TNFAIP3 is required for FGFR1 activation-promoted proliferation and tumorigenesis of premalignant DCIS.COM human mammary epithelial cells.
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TNFAIP3 是 FGFR1 激活促进癌前 DCIS.COM 人乳腺上皮细胞增殖和肿瘤发生所必需的
DOI：
10.1186/s13058-018-1024-9
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发表时间：
2018-08-15
期刊：
影响因子：
--
通讯作者：
He T
中科院分区：
文献类型：
--
作者：
Yang M;Yu X;Li X;Luo B;Yang W;Lin Y;Li D;Gan Z;Xu J;He T
尽管导管原位癌 (DCIS) 是一种非浸润性乳腺癌，但许多 DCIS 病变可能进展为浸润性癌症，并且导致其进展的基因和途径很大程度上未知。 FGFR1在细胞增殖、分化和癌变中发挥重要作用。本研究的目的是在非侵入性 DCIS 模型中检查 FGFR1 信号传导在基因表达、细胞增殖、肿瘤生长和进展中的作用。用空载体转染 DCIS.COM 细胞以生成 DCIS-Ctrl 细胞。用 AP20187 诱导型 iFGFR1 载体转染 DCIS-iFGFR1 细胞，生成 DCIS-iFGFR1 细胞。 iFGFR1 由 v-Src 肉豆蔻酰化膜靶向序列、FGFR1 胞质结构域和 AP20187 诱导的 FKBP12 二聚化结构域组成，可模拟 FGFR1 信号传导。采用 CRISPR/Cas9 系统敲除 DCIS-iFGFR1 细胞中的 ERK1、ERK2 或 TNFAIP3。使用/不使用 AP20187 以及使用/不使用 FGFR1、MEK 或 ERK1/2 抑制剂处理建立的细胞系。这些治疗的效果通过蛋白质印迹、RNA-Seq、实时 RT-PCR、细胞增殖、微球生长、异种移植肿瘤生长和肿瘤组织病理学测定来确定。 DCIS-iFGFR1 细胞中 iFGFR1 信号的激活增强了 ERK1/2 活性，诱导部分上皮间质转化 (EMT) 并增加细胞增殖。 iFGFR1 信号传导的激活促进 DCIS 生长并进展为源自小鼠 DCIS-iFGFR1 细胞的侵袭性癌症。 iFGFR1 信号传导的激活还改变了涉及细胞增殖、迁移、癌症途径以及其他分子和细胞功能的 946 个基因的表达水平。 TNFAIP3 是一种泛素编辑酶，通过 FGFR1 激酶活性和 ERK2 依赖性方式的 iFGFR1 信号传导上调。重要的是，TNFAIP3敲除不仅抑制了AP20187诱导的DCIS-iFGFR1细胞的增殖和肿瘤生长，而且还进一步降低了未经AP20187治疗的DCIS-iFGFR1细胞的基线增殖和肿瘤生长。 iFGFR1 的激活可促进 ERK1/2 活性、EMT、细胞增殖、肿瘤生长、DCIS 进展为浸润性癌症，并改变 DCIS-iFGFR1 细胞的基因表达谱。 iFGFR1 的激活以 ERK2 依赖性方式上调 TNFAIP3，并且 TNFAIP3 是 iFGFR1 激活促进的 DCIS.COM 细胞增殖、乳腺球生长、肿瘤生长和进展所必需的。这些结果表明 TNFAIP3 可能是抑制 FGFR1 信号传导促进的 DCIS 生长和进展的潜在靶点。本文的在线版本 (10.1186/s13058-018-1024-9) 包含补充材料，可供授权用户使用。
Although ductal carcinoma in situ (DCIS) is a non-invasive breast cancer, many DCIS lesions may progress to invasive cancer and the genes and pathways responsible for its progression are largely unknown. FGFR1 plays an important role in cell proliferation, differentiation and carcinogenesis. The purpose of this study is to examine the roles of FGFR1 signaling in gene expression, cell proliferation, tumor growth and progression in a non-invasive DCIS model. DCIS.COM cells were transfected with an empty vector to generate DCIS-Ctrl cells. DCIS-iFGFR1 cells were transfected with an AP20187-inducible iFGFR1 vector to generate DCIS-iFGFR1 cells. iFGFR1 consists of the v-Src myristoylation membrane-targeting sequence, FGFR1 cytoplasmic domain and the AP20187-inducible FKBP12 dimerization domain, which simulates FGFR1 signaling. The CRISPR/Cas9 system was employed to knockout ERK1, ERK2 or TNFAIP3 in DCIS-iFGFR1 cells. Established cell lines were treated with/without AP20187 and with/without FGFR1, MEK, or ERK1/2 inhibitor. The effects of these treatments were determined by Western blot, RNA-Seq, real-time RT-PCR, cell proliferation, mammosphere growth, xenograft tumor growth, and tumor histopathological assays. Activation of iFGFR1 signaling in DCIS-iFGFR1 cells enhanced ERK1/2 activities, induced partial epithelial-to-mesenchymal transition (EMT) and increased cell proliferation. Activation of iFGFR1 signaling promoted DCIS growth and progression to invasive cancer derived from DCIS-iFGFR1 cells in mice. Activation of iFGFR1 signaling also altered expression levels of 946 genes involved in cell proliferation, migration, cancer pathways, and other molecular and cellular functions. TNFAIP3, a ubiquitin-editing enzyme, is upregulated by iFGFR1 signaling in a FGFR1 kinase activity and in an ERK2-dependent manner. Importantly, TNFAIP3 knockout not only inhibited the AP20187-induced proliferation and tumor growth of DCIS-iFGFR1 cells, but also further reduced baseline proliferation and tumor growth of DCIS-iFGFR1 cells without AP20187 treatment. Activation of iFGFR1 promotes ERK1/2 activity, EMT, cell proliferation, tumor growth, DCIS progression to invasive cancer, and altered the gene expression profile of DCIS-iFGFR1 cells. Activation of iFGFR1 upregulated TNFAIP3 in an ERK2-dependent manner and TNFAIP3 is required for iFGFR1 activation-promoted DCIS.COM cell proliferation, mammosphere growth, tumor growth and progression. These results suggest that TNFAIP3 may be a potential target for inhibiting DCIS growth and progression promoted by FGFR1 signaling. The online version of this article (10.1186/s13058-018-1024-9) contains supplementary material, which is available to authorized users.
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DOI：
10.1038/nrm3528
发表时间：
2013-03
期刊：
Nature reviews. Molecular cell biology
影响因子：
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50.3
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DOI：
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发表时间：
2017-10-15
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
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DOI：
10.1016/j.apsb.2014.02.003
发表时间：
2014-06
期刊：
Acta pharmaceutica Sinica. B
影响因子：
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