Department of Obstetrics and Gynecology, Section of Gynecologic Oncology, University of Chicago, Chicago, IL 60637, United States of America.
Department of Pathology, University of Chicago, Chicago, IL 60637, United States of America.
Gynecol Oncol. 2019 May;153(2):405-415. doi: 10.1016/j.ygyno.2019.01.020. Epub 2019 Feb 20.
Ovarian cancer (OvCa) metastasis requires the coordinated motility of both cancer and stromal cells. Cellular movement is a dynamic process that involves the synchronized assembly of f-actin bundles into cytoskeletal protrusions by fascin. Fascin directly binds f-actin and is an integral component of filopodia, lamellapodia and stress fibers. Here, we examine the expression pattern and function of fascin in the cancer and stromal cells of OvCa tumors.
Fascin expression was evaluated in human cells and tissues using immunohistochemistry and immunofluorescence. The functional role of fascin in cancer and stromal cells was assessed with in vitro functional assays, an ex vivo colonization assay and in vivo metastasis assays using siRNA/shRNA and an inhibitor. The effect of fascin inhibition on Cdc42 and Rac1 activity was evaluated using GTPase activity assays and immunofluorescence.
Fascin expression was found to be higher in the stromal cell, when compared to the cancer cell, compartment of ovarian tumors. The low expression of fascin in the cancer cells of the primary tumor indicated a favorable prognosis for non-serous OvCa patients. In vitro, both knockdown and pharmacologic inhibition of fascin decreased the migration of cancer and stromal cells. The inhibition of fascin impaired Cdc42 and Rac1 activity in cancer cells, and cytoskeletal reorganization in the cancer and stromal cells. Inhibition of fascin ex vivo blocked OvCa cell colonization of human omental tissue and in vivo prevented and reduced OvCa metastases in mice. Likewise, knockdown of fascin specifically in the OvCa cells using a fascin-specific lentiviral-shRNA also blocked metastasis in vivo.
This study reveals the therapeutic potential of pharmacologically inhibiting fascin in both cancer and stromal cells of the OvCa tumor microenvironment.
卵巢癌(OvCa)转移需要癌细胞和基质细胞的协调运动。细胞运动是一个动态过程,涉及到 fascin 对 f-肌动蛋白束的同步组装,形成细胞骨架突起。fascin 直接结合 f-肌动蛋白,是丝状伪足、片状伪足和应激纤维的组成部分。在这里,我们研究了 fascin 在 OvCa 肿瘤的癌细胞和基质细胞中的表达模式和功能。
使用免疫组织化学和免疫荧光法评估 fascin 在人细胞和组织中的表达。使用体外功能测定、体外定植测定和体内转移测定,评估 fascin 在癌细胞和基质细胞中的功能作用,使用 siRNA/shRNA 和抑制剂。使用 GTPase 活性测定和免疫荧光法评估 fascin 抑制对 Cdc42 和 Rac1 活性的影响。
发现卵巢肿瘤的基质细胞中 fascin 的表达高于癌细胞。原发肿瘤中 fascin 的低表达表明非浆液性 OvCa 患者的预后较好。在体外,fascin 的敲低和药理学抑制均降低了癌细胞和基质细胞的迁移。fascin 的抑制抑制了癌细胞中 Cdc42 和 Rac1 的活性,并影响了癌细胞和基质细胞的细胞骨架重组。体外抑制 fascin 阻断了 OvCa 细胞对人网膜组织的定植,体内抑制 fascin 阻止并减少了小鼠 OvCa 转移。同样,使用 fascin 特异性慢病毒-shRNA 特异性敲低 OvCa 细胞中的 fascin 也阻止了体内转移。
这项研究揭示了在 OvCa 肿瘤微环境中的癌细胞和基质细胞中,药理学抑制 fascin 的治疗潜力。