Department of Clinical Laboratory, Beijing Shijitan Hospital, Capital Medical University, 10 Tieyi Road, Haidian District, Beijing, 100038, China.
Beijing Key Laboratory of Urinary Cellular Molecular Diagnostics, Beijing, 100038, China.
Invest New Drugs. 2019 Oct;37(5):935-947. doi: 10.1007/s10637-018-0709-3. Epub 2019 Jan 4.
Prostate-specific antigen (PSA) has been widely used as the unique serum biomarker for the diagnosis of prostate cancer (PCa). When PSA is moderately increased (e.g., 4-10 ng/ml), it is difficult to differentiate benign prostatic hyperplasia (BPH) from cancer. The diagnostic test (i.e., prostate biopsy) is invasive, adding pain and economic burden to the patient. Urine samples are more convenient, non-invasive and readily available than blood. We sought to determine whether ferritin might be the potential urinary biomarker in prostate cancer diagnosis. Using two-dimensional electrophoresis (2DE) followed by mass spectrometry (MS), differentially expressed urinary proteins among patients with PCa, BPH and normal controls were obtained. The ferritin heavy chain (FTH) gene, ferritin light chain (FTL) gene and protein expression of BPH-1 cells and PC-3 cells were analyzed by real-time quantitative PCR and Western blotting, respectively. Stable FTH or FTL silenced cell lines were generated by small hairpin(sh) RNA lentiviral transfection. The function of the cell lines was evaluated by the colony formation assay, transwell assay, and flow cytometry. Compared with BPH and normal controls, 15 overexpressed proteins, including FTH and FTL, were identified in the urine of the PCa group. FTH and FTL were also highly expressed in PC-3 cell lines compared with BPH-1 cells. FTH-silenced cells showed reduced cell proliferation, migration and increased cell apoptosis. FTL-silenced cells showed increased proliferation and migration abilities. There are differences in urinary proteins among patients with PCa, BPH and normal controls. FTH and FTL play different roles in PCa cells and are potential biomarkers for PCa.
前列腺特异性抗原 (PSA) 已被广泛用作前列腺癌 (PCa) 诊断的唯一血清生物标志物。当 PSA 中度升高（例如，4-10ng/ml）时，很难将良性前列腺增生 (BPH) 与癌症区分开来。诊断性检查（即前列腺活检）是有创的，会给患者带来疼痛和经济负担。尿液样本比血液更方便、无创且易于获得。我们试图确定铁蛋白是否可能成为前列腺癌诊断的潜在尿液生物标志物。使用二维电泳 (2DE) 结合质谱 (MS)，获得了 PCa、BPH 和正常对照组患者之间差异表达的尿液蛋白。通过实时定量 PCR 和 Western blot 分别分析了铁蛋白重链 (FTH) 基因、铁蛋白轻链 (FTL) 基因和 BPH-1 细胞和 PC-3 细胞的蛋白表达。通过小发夹(sh)RNA 慢病毒转染生成稳定的 FTH 或 FTL 沉默细胞系。通过集落形成试验、Transwell 试验和流式细胞术评估细胞系的功能。与 BPH 和正常对照组相比，在 PCa 组的尿液中鉴定出 15 种过表达蛋白，包括 FTH 和 FTL。与 BPH-1 细胞相比，PC-3 细胞系中 FTH 和 FTL 的表达也更高。沉默 FTH 的细胞显示出细胞增殖、迁移减少和细胞凋亡增加。沉默 FTL 的细胞显示出增殖和迁移能力增强。PCa、BPH 和正常对照组患者的尿液蛋白存在差异。FTH 和 FTL 在 PCa 细胞中发挥不同的作用，是 PCa 的潜在生物标志物。
