一种靶向WASF3–CYFIP1复合体的“订书钉”肽可抑制癌细胞侵袭选编.pptx

Targeting the WASF3–CYFIP1 Complex Using Stapled Peptides Suppresses Cancer Cell Invasion 一种靶向WASF3–CYFIP1复合体的“订书钉”肽可抑制癌细胞侵袭 ;;1. Introduction;WASF3的基因可促进细胞移动、侵袭和新陈代谢，在高度侵袭性的乳腺癌和前列腺癌细胞中可高水平的持续性表达。 1、调节 KISS1, ZEB1, 及 miRNA-200s等蛋白 2、被JAK2, HSP70, ABL, 及 HIF1蛋白调节 3、与 ATAD3A线粒体蛋白相互作用从而调节它在线粒体膜上的稳定性 WASF3与其它蛋白相互作用形成CYFIP1–NCKAP1二聚体(主要)或ABI2–HSPC300–WASF三聚体发挥作用，;“stapled peptides”： WASF Helix Mimics (WAHM) 优点：化学结构稳定，细胞透性强，抗蛋白酶降解 缺点：近来才被应用于生物治疗学方面，仍面临挑战，但有些研究已进入I期临床 试验阶段。研究人员证明stapled peptides可阻断WASF3与CYFIP1之间的相互作用从而能够抑制WASF3的功能。;2. Materials and Methods;Cell lines : MDA-MB-231 cells, DU145 cells standard assays: Lentiviral transduction, cell proliferation assays, wound-healing assays, Transwell invasion assays, Western blotting, flow cytometry, and real-time RT-PCR analysis;3.1. Loss of CYFIP1 protein leads to WASF3 instability, resulting in suppression of invasion;These data suggest that disrupting the protein–protein complex involving CYFIP1 could lead to suppression of invasion by affecting WASF3 function.;3.2. Role of WASF family members in invasion and metastasis;These observations suggest that, although the WASF proteins have been implicated in cell movement, only WASF3 is particularly and specifically associated with invasion and metastasis of cancer cells.;3.3. Targeting the CYFIP1–WASF interaction with stapled peptides;To determine whether the stapled peptides formed a complex with WASF3 within the cells, MDA-MB-231 cells were exposed to biotinylated WAHM1 and WAHM2 for 6 hours, after which the cells were lysed and pulldown assays were performed using avidin-coated beads.;3.4. WAHM1/2 peptides suppress cancer cell motility and invasion;Thus, targeting the CYFIP1–WASF3 interaction with stapled peptides affects cancer cell cytoskeleton organization, motility, and invasion, consistent with knockdown of WASF3 function achieved by other means.;ShRNA knockdown of CYFIP1 leads to WASF3 protein destabilization (Fig. 1). In the stapled peptide treated cells, however