Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada.
J Proteome Res. 2009 Oct;8(10):4705-13. doi: 10.1021/pr900411g.
Ovarian cancer remains a deadly threat to women as the disease is often diagnosed in the late stages when the chance of survival is low. There are no good biomarkers available for early detection and only a few markers have shown clinical utility for prognosis, response to therapy and disease recurrence. We mined conditioned media of four ovarian cancer cell lines (HTB75, TOV-112D, TOV-21G and RMUG-S) by two-dimensional liquid chromatography-mass spectrometry. Each cell line represented one of the major histological types of epithelial ovarian cancer. We identified 2039 proteins from which 228 were extracellular and 192 were plasma membrane proteins. Within the latter list, we identified several known markers of ovarian cancer including three that are well established, namely, CA-125, HE4, and KLK6. The list of 420 extracellular and membrane proteins was cross-referenced with the proteome of ascites fluid to generate a shorter list of 51 potential biomarker candidates. According to Ingenuity Pathway Analysis, two of the top 10 diseases associated with the list of 51 proteins were cancer and reproductive diseases. We selected nine proteins for preliminary validation using 20 serum samples from healthy women and 10 from women with ovarian cancer. Of the nine proteins, clusterin (increase) and IGFBP6 (decrease) showed significant differences between women with or without ovarian cancer. We conclude that in-depth proteomic analysis of cell culture supernatants of ovarian cancer cell lines can identify potential ovarian cancer biomarkers that are worth further clinical validation.
卵巢癌仍然是女性的致命威胁，因为这种疾病通常在晚期诊断，此时生存机会较低。目前尚无用于早期检测的良好生物标志物，只有少数标志物在预后、对治疗的反应和疾病复发方面显示出临床实用性。我们通过二维液相色谱-质谱法对四种卵巢癌细胞系（HTB75、TOV-112D、TOV-21G 和 RMUG-S）的条件培养基进行了挖掘。每个细胞系代表上皮性卵巢癌的一种主要组织学类型。我们从 2039 种蛋白质中鉴定出 228 种细胞外蛋白质和 192 种质膜蛋白质。在后一组列表中，我们鉴定出了几种已知的卵巢癌标志物，包括三种已确立的标志物，即 CA-125、HE4 和 KLK6。420 种细胞外和膜蛋白的列表与腹水的蛋白质组进行了交叉引用，生成了 51 种潜在生物标志物候选物的更短列表。根据 Ingenuity 通路分析，与这 51 种蛋白质列表相关的前 10 种疾病中有两种是癌症和生殖系统疾病。我们选择了 9 种蛋白质，使用 20 份来自健康女性和 10 份来自卵巢癌女性的血清样本进行初步验证。在这 9 种蛋白质中，簇蛋白（增加）和 IGFBP6（减少）在有无卵巢癌的女性之间存在显著差异。我们得出结论，深入分析卵巢癌细胞系的细胞培养上清液中的蛋白质组可以鉴定出潜在的卵巢癌生物标志物，值得进一步的临床验证。
