Microparticles From Ovarian Carcinomas Are Shed Into Ascites and Promote Cell Migration
Microparticles From Ovarian Carcinomas Are Shed Into Ascites and Promote Cell Migration
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DOI：
10.1097/igc.0b013e318241d9b9
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发表时间：
2012-05-01
影响因子：
4.8
通讯作者：
Swisher, Elizabeth M.
中科院分区：
文献类型：
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作者：
Press, Joshua Z.;Reyes, Morayma;Swisher, Elizabeth M.
目的：微粒是由细胞膜成分组成的细胞衍生囊泡（0.5-1.0 μ m），其从各种细胞（包括上皮细胞）表面主动脱落。我们比较了卵巢癌和良性卵巢病变的妇女之间的腹水中的微粒，并分离出肿瘤衍生的（上皮细胞粘附分子[EpCAM]阳性）微粒的功能分析和蛋白质组学。材料和方法：病例包括8例良性卵巢肿瘤和41例卵巢癌。来自高级别III期浆液性癌的腹水用于功能和蛋白质组学分析。使用EpCAM包被的珠分离癌细胞，通过超离心/流式细胞术分离微粒，并使用标记物（例如EpCAM）实现分选。用3种卵巢癌细胞系进行结合和迁移测定。结果：良性盆腔积液中的微粒与早期和晚期腹水中的微粒相似（2.4 vs 2.8 vs 2.0 x 10（6）微粒/mL）。晚期患者EpCAM阳性微粒的比例高于早期或良性疾病（13.3% vs 2.5% vs 2.1%; P = 0.001），浆液性组织学检查的EpCAM阳性微粒的比例高于类浆液性组织学检查（13.2% vs 1.8%; P = 0.01）。微粒结合到3种培养的细胞系的表面，并内化到EpCAM阳性微粒中，导致比单独的缓冲液或EpCAM阴性微粒更多的细胞迁移（P = 0.007）。随着EpCAM阳性微粒数量的增加，观察到剂量依赖性增加。蛋白质组学研究表明，EPCAM阳性微粒中的大多数蛋白质与癌细胞共享，并且许多蛋白质与细胞运动和侵袭有关，如纤连蛋白、细丝蛋白A、波形蛋白、肌球蛋白9和纤维蛋白原fibrin.Conclusions：晚期和浆液性卵巢癌的腹水中含有大量的肿瘤来源的微粒。在体外，这些微粒与癌细胞结合并刺激迁移。腹水中肿瘤来源的微粒可能介导浆液性卵巢癌腹膜扩散的倾向。
Objective: Microparticles are cellular-derived vesicles (0.5-1.0 mu m) composed of cell membrane components, which are actively shed from the surface of various cells, including epithelial cells. We compared microparticles in ascites between women with ovarian carcinoma and women with benign ovarian pathology, and isolated tumor-derived (epithelial cell adhesion molecule [EpCAM]-positive) microparticles for functional analysis and proteomics.Materials and Methods: Cases included 8 patients with benign ovarian neoplasms and 41 with ovarian carcinoma. Ascites from a high-grade stage III serous carcinoma was used for functional and proteomic analysis. Cancer cells were isolated using EpCAM-coated beads, microparticles were isolated by ultracentrifugation/flow cytometry, and sorting was achieved using markers (eg, EpCAM). Binding and migrations assays were performed with 3 ovarian cancer cell lines. Proteomic analysis of EpCAM-positive microparticles and ascites cancer cells was performed by mass spectrometry.Results: Microparticles in benign pelvic fluid were similar to early and advanced-stage ascites (2.4 vs 2.8 vs 2.0 x 10(6) microparticles/mL). Advanced stage had a greater proportion of EpCAM-positive microparticles than early or benign disease (13.3% vs 2.5% vs 2.1%; P = 0.001), and serous histology had more than endometrioid (13.2% vs 1.8%; P = 0.01). Microparticles bound to the surface of 3 cultured cell lines, and were internalized into the EpCAM-positive microparticles, resulting in more cell migration than buffer alone or EpCAM-negative microparticles (P = 0.007). A dose-dependent increase was seen with increasing numbers of EpCAM-positive microparticles. Proteomics revealed that most proteins in EPCAM-positive microparticles were shared with cancer cells, and many are associated with cell motility and invasion, such as fibronectin, filamin A, vimentin, myosin-9, and fibrinogen.Conclusions: Ascites from advanced-stage and serous ovarian carcinomas contain large numbers of tumor-derived microparticles. In vitro, these microparticles bind to cancer cells and stimulate migration. Tumor-derived microparticles in ascites could mediate the predilection for peritoneal spread in serous ovarian carcinomas.