P300/CBP-associated factor (PCAF)-mediated acetylation of Fascin at lysine 471 inhibits its actin-bundling activity and tumor metastasis in esophageal cancer.
P300/CBP-associated factor (PCAF)-mediated acetylation of Fascin at lysine 471 inhibits its actin-bundling activity and tumor metastasis in esophageal cancer.
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P300/CBP 相关因子 (PCAF) 介导的 Fascin 471 位赖氨酸乙酰化可抑制其肌动蛋白成束活性和食管癌肿瘤转移
DOI：
10.1002/cac2.12221
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发表时间：
2021-12
期刊：
影响因子：
--
通讯作者：
Li EM
中科院分区：
文献类型：
--
作者：
Cheng YW;Zeng FM;Li DJ;Wang SH;He JZ;Guo ZC;Nie PJ;Wu ZY;Shi WQ;Wen B;Xu XE;Liao LD;Li ZM;Wu JY;Zhan J;Zhang HQ;Chang ZJ;Zhang K;Xu LY;Li EM
Fascin对于癌细胞丝状伪足形成和肿瘤转移至关重要，并且在功能上受到翻译后修饰的调节。然而，Fascin是否以及如何通过乙酰化调节仍不清楚。本研究探讨Fascin乙酰化的调控及其在丝状伪足形成和肿瘤转移中的作用。采用免疫沉淀和谷胱甘肽-S-转移酶下拉试验检测Fascin与乙酰转移酶P300/CBP-相关因子（PCAF）之间的相互作用，并采用免疫荧光法研究它们的共定位。进行体外乙酰化测定以通过使用质谱法鉴定肌成束蛋白乙酰化位点。生成针对乙酰化Fascin的特异性抗体，并使用蛋白质印迹法通过过表达和敲低PCAF表达来检测食管鳞状细胞癌（ESCC）细胞中PCAF介导的Fascin乙酰化。进行体外细胞迁移试验，并建立异种移植模型来研究体内肿瘤转移。使用活细胞成像和光漂白后的荧光恢复来评估乙酰化Fascin在丝状伪足形成中的功能和动力学。应用免疫组化方法评价乙酰化Fascin和PCAF在食管鳞癌中的表达及其临床意义。Fascin与PCAF直接相互作用并共定位于细胞质中，并被PCAF在赖氨酸471（K471）处乙酰化。使用特异性抗AcK 471-Fascin抗体，发现Fascin在ESCC细胞中被乙酰化，并且乙酰化水平因此在PCAF过表达后增加，并且在PCAF敲低后降低。在功能上，Fascin-K471乙酰化显著抑制体外ESCC细胞迁移和体内肿瘤转移，而Fascin-K471去乙酰化表现出有效的致癌功能。此外，Fascin-K471乙酰化降低了丝状伪足的长度和密度，以及ESCC细胞的寿命，而其去乙酰化产生了相反的效果。在丝状体中，K471-乙酰化的Fascin显示出快速的动态交换，这表明由于其减弱的肌动蛋白捆绑活性，它仍然保持其单体形式。临床上，ESCC组织中高水平的AcK 471 ‐Fascin与ESCC患者的总生存期和无病生存期延长密切相关。成束蛋白直接与PCAF相互作用，并在ESCC细胞中在赖氨酸471处乙酰化。Fascin-K471乙酰化通过损害其肌动蛋白捆绑活性减少丝状伪足形成来抑制ESCC细胞迁移和肿瘤转移。
Fascin is crucial for cancer cell filopodium formation and tumor metastasis, and is functionally regulated by post‐translational modifications. However, whether and how Fascin is regulated by acetylation remains unclear. This study explored the regulation of Fascin acetylation and its corresponding roles in filopodium formation and tumor metastasis. Immunoprecipitation and glutathione‐S‐transferase pull‐down assays were performed to examine the interaction between Fascin and acetyltransferase P300/CBP‐associated factor (PCAF), and immunofluorescence was used to investigate their colocalization. An in vitro acetylation assay was performed to identify Fascin acetylation sites by using mass spectrometry. A specific antibody against acetylated Fascin was generated and used to detect the PCAF‐mediated Fascin acetylation in esophageal squamous cell carcinoma (ESCC) cells using Western blotting by overexpressing and knocking down PCAF expression. An in vitro cell migration assay was performed, and a xenograft model was established to study in vivo tumor metastasis. Live‐cell imaging and fluorescence recovery after photobleaching were used to evaluate the function and dynamics of acetylated Fascin in filopodium formation. The clinical significance of acetylated Fascin and PCAF in ESCC was evaluated using immunohistochemistry. Fascin directly interacted and colocalized with PCAF in the cytoplasm and was acetylated at lysine 471 (K471) by PCAF. Using the specific anti‐AcK471‐Fascin antibody, Fascin was found to be acetylated in ESCC cells, and the acetylation level was consequently increased after PCAF overexpression and decreased after PCAF knockdown. Functionally, Fascin‐K471 acetylation markedly suppressed in vitro ESCC cell migration and in vivo tumor metastasis, whereas Fascin‐K471 deacetylation exhibited a potent oncogenic function. Moreover, Fascin‐K471 acetylation reduced filopodial length and density, and lifespan of ESCC cells, while its deacetylation produced the opposite effect. In the filipodium shaft, K471‐acetylated Fascin displayed rapid dynamic exchange, suggesting that it remained in its monomeric form owing to its weakened actin‐bundling activity. Clinically, high levels of AcK471‐Fascin in ESCC tissues were strongly associated with prolonged overall survival and disease‐free survival of ESCC patients. Fascin interacts directly with PCAF and is acetylated at lysine 471 in ESCC cells. Fascin‐K471 acetylation suppressed ESCC cell migration and tumor metastasis by reducing filopodium formation through the impairment of its actin‐bundling activity.
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影响因子：
56.9
作者：
Choudhary, Chunaram;Kumar, Chanchal;Mann, Matthias
通讯作者：
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影响因子：
16.6
作者：
Fournier, Marjorie;Orpinell, Meritxell;Grauffel, Cedric;Scheer, Elisabeth;Garnier, Jean-Marie;Ye, Tao;Chavant, Virginie;Joint, Mathilde;Esashi, Fumiko;Dejaegere, Annick;Gonczy, Pierre;Tora, Laszlo
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影响因子：
3.3
作者：
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通讯作者：
Adams, Josephine C.
影响因子：
7.5
作者：
Jacquemet, Guillaume;Hamidi, Hellyeh;Ivaska, Johanna
通讯作者：
Ivaska, Johanna
影响因子：
4.1
作者：
Kuninger, David;Lundblad, James;Rotwein, Peter
通讯作者：
Rotwein, Peter