Fenretinide Perturbs Focal Adhesion Kinase in Premalignant and Malignant Human Oral Keratinocytes. Fenretinide's Chemopreventive Mechanisms Include ECM Interactions.
Fenretinide Perturbs Focal Adhesion Kinase in Premalignant and Malignant Human Oral Keratinocytes. Fenretinide's Chemopreventive Mechanisms Include ECM Interactions.
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DOI：
10.1158/1940-6207.capr-14-0418
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发表时间：
2015-05
期刊：
影响因子：
--
通讯作者：
Mallery SR
中科院分区：
文献类型：
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作者：
Han BB;Li S;Tong M;Holpuch AS;Spinney R;Wang D;Border MB;Liu Z;Sarode S;Pei P;Schwendeman SP;Mallery SR
膜相关蛋白，粘着斑激酶(FAK)，调节细胞-细胞外基质的相互作用，也传递促进生存和增殖的信号。值得注意的是，上皮内FAK水平升高伴随着癌前口腔上皮内瘤变(OIN)向口腔鳞状细胞癌(OSCC)的转化。OIN化学预防是一种以患者为中心的最佳策略，可防止口腔鳞癌的合并症和死亡率。化学预防癌症的合成维生素A衍生物芬维甲素已显示出与蛋白质结合的能力，例如mTOR和视黄醇结合蛋白相互作用。这些研究使用了连续的人类口腔角质形成细胞(正常-HPVE6/E7-转导的口腔鳞状细胞癌)来评估潜在的非维甲酸-FAK药物蛋白相互作用以及对细胞生长调节和运动的功能后果。分子模拟研究表明，Fenretinide的结合亲和力比FAK的激酶结构域上的天然配体(ATP)高约200倍。Fenretinide还在FAK的FERM结构域显示中间结合，并与最接近的FAK类似物Pyk2的ATP结合位点相互作用。芬维甲素通过诱导细胞凋亡和G2/M期阻滞而显著抑制细胞增殖。Fenretinide处理的细胞还显示了F-肌动蛋白的破坏，显著抑制了合成基底膜的定向迁移和侵袭，并减少了促生长因子的磷酸化。一种商业化的FAK抑制剂不能抑制细胞的侵袭。值得注意的是，虽然FAK的FERM结构域指导细胞侵袭，但FAK抑制剂的靶向是激酶结构域。此外，FAK特异性siRNA处理的细胞显示出中等的细胞迁移能力；这一数据表明，已建立的促进迁移的Pyk2共同作用。我们的数据表明，非维甲酸是唯一能够阻断FAK和PYK2的S促生存和提高运动能力的作用，并进一步将非维甲酸的化学预防作用扩展到诱导细胞凋亡和分化之外。
The membrane-associated protein, focal adhesion kinase (FAK), modulates cell-extracellular matrix interactions and also conveys pro-survival and proliferative signals. Notably, increased intraepithelial FAK levels accompany transformation of premalignant oral intraepithelial neoplasia (OIN) to oral squamous cell carcinoma (OSCC). OIN chemoprevention is a patient-centric, optimal strategy to prevent OSCC’s co-morbidities and mortality. The cancer chemopreventive and synthetic vitamin A derivative, fenretinide, has demonstrated protein-binding capacities e.g. mTOR and retinol binding protein interactions. These studies employed a continuum of human oral keratinocytes (normal-HPV E6/E7-transduced-OSCC) to assess potential fenretinide-FAK drug protein interactions and functional consequences on cellular growth regulation and motility. Molecular modeling studies demonstrated fenretinide has ~200-fold greater binding affinity relative to the natural ligand (ATP) at FAK’s kinase domain. Fenretinide also shows intermediate binding at FAK’s FERM domain and interacts at the ATP-binding site of the closest FAK analogue, Pyk2. Fenretinide significantly suppressed proliferation via induction of apoptosis and G2/M cell cycle blockade. Fenretinide-treated cells also demonstrated F-actin disruption, significant inhibition of both directed migration and invasion of a synthetic basement membrane, and decreased phosphorylation of growth-promoting kinases. A commercially available FAK inhibitor did not suppress cell invasion. Notably, while FAK’s FERM domain directs cell invasion, FAK inhibitors target the kinase domain. In addition, FAK-specific siRNA treated cells showed an intermediate cell migration capacity; data which suggest co-contribution of the established migrating-enhancing Pyk2. Our data imply that fenretinide is uniquely capable of disrupting FAK’s and Pyk2’s pro-survival and mobility-enhancing effects and further extend fenretinide’s chemopreventive contributions beyond induction of apoptosis and differentiation.