Municipal Key Laboratory for Diseases Related to Women's Reproductive and Endocrine Systems, Obstetrics and Gynecology Hospital, Fudan University, Shanghai 200011, People's Republic of China.
J Proteome Res. 2010 Dec 3;9(12):6180-90. doi: 10.1021/pr100822v. Epub 2010 Oct 28.
Peritoneal implantation is the most common metastatic pattern of epithelial ovarian cancer, and the five-year survival rate of patients is dramatically decreased when large-scale peritoneal metastasis occurs. This study aimed to determine serum proteins that could be used to detect early peritoneal metastasis of ovarian cancer. The secreted (or shed) proteins of the ovarian cancer cell line SKOV-3 were analyzed using LC-MS/MS, and 97 proteins were identified in the SKOV-3 culture supernatant. After the SKOV-3 cells were xenografted into the peritoneal cavities of nude mice, 3 of the 97 proteins were detected in animal sera. Following enzyme-linked immunosorbent assay (ELISA)-based screening of clinical blood samples, one of the three proteins, 14-3-3 zeta, was identified as a candidate biomarker. The average serum levels of 14-3-3 zeta in patients with epithelial ovarian cancer and benign gynecological diseases were significantly different. The expression of 14-3-3 zeta was associated with the degree of cancer peritoneal metastasis, the emergence of ascites, bilateral involvement, and the clinical stage and substage. Using 14-3-3 zeta, the overall diagnostic accuracy for ovarian cancer was greatly improved. Furthermore, siRNA-based experiments demonstrated that 14-3-3 zeta was responsible for approximately 62, 65, and 30% of the migratory, invasive, and implantation abilities of SKOV-3 cells, respectively. The present results demonstrated that the nude mouse xenograft model is an efficient system for performing function-oriented biomarker discovery, which can be used for a variety of research tasks in future molecular diagnoses, targeted therapies, and ovarian cancer vaccine development.
腹膜种植是上皮性卵巢癌最常见的转移方式，当发生大规模腹膜转移时，患者的五年生存率显著降低。本研究旨在确定可用于检测卵巢癌早期腹膜转移的血清蛋白。使用 LC-MS/MS 分析卵巢癌细胞系 SKOV-3 的分泌蛋白（或脱落蛋白），在 SKOV-3 培养上清液中鉴定出 97 种蛋白质。将 SKOV-3 细胞异种移植到裸鼠腹腔后，在动物血清中检测到 97 种蛋白质中的 3 种。通过基于酶联免疫吸附试验（ELISA）的临床血液样本筛选后，发现这 3 种蛋白质中的 14-3-3 zeta 可作为候选生物标志物。上皮性卵巢癌和良性妇科疾病患者的血清 14-3-3 zeta 平均水平有显著差异。14-3-3 zeta 的表达与癌症腹膜转移程度、腹水出现、双侧受累以及临床分期和亚分期有关。使用 14-3-3 zeta，大大提高了卵巢癌的整体诊断准确性。此外，基于 siRNA 的实验表明，14-3-3 zeta 分别负责 SKOV-3 细胞约 62%、65%和 30%的迁移、侵袭和植入能力。本研究结果表明，裸鼠异种移植模型是一种进行面向功能的生物标志物发现的有效系统，可用于未来分子诊断、靶向治疗和卵巢癌疫苗开发的各种研究任务。
