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应用特异性单克隆抗体检测膀胱癌和肾细胞癌中的生存素表达。

Detection of survivin expression in bladder cancer and renal cell carcinoma using specific monoclonal antibodies.

机构信息

Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Clinical Laboratory, Peking University School of Oncology, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China.

出版信息

Oncol Rep. 2018 Jun;39(6):2817-2828. doi: 10.3892/or.2018.6359. Epub 2018 Apr 5.

Abstract

Survivin, which is highly expressed in the majority of tumors, but not in most normal adult tissues, has been identified to have significant clinical applications. In the present study, using survivin‑specific monoclonal antibodies (mAbs), we aimed to establish methods for detecting the expression of survivin in cancer cell lines, serum samples, urine samples and cancer tissues from patients with bladder cancer (BCa) and renal cell carcinoma (RCC), and to evaluate the efficacy of survivin as a tumor marker in the surveillance of BCa and RCC. First, mAbs were labeled with horseradish peroxidase (HRP), and a sandwich enzyme‑linked immunosorbent assay (ELISA) with mAbs and HRP‑conjugated mAbs was developed to detect survivin expression in serum and urine samples from BCa and RCC patients, with samples from healthy controls (HCs) used for comparison. The HRP‑conjugated mAbs were also used to detect survivin expression in cancer cell lines by western blotting. Survivin expression in cancer tissues from BCa patients was also evaluated by immunohistochemistry. The results showed that the sandwich ELISA was successfully established, and significantly higher expression of survivin was subsequently detected in BCa and RCC patients as compared with HCs in both urinary and serum samples (P<0.05), and was more pronounced in urine. The HRP‑mAbs could recognize survivin in cancer cell lines. Western blotting and immunohistochemistry results confirmed survivin expression in the 5637 BCa cell line, as well as BCa tissues. In addition, the expressions of survivin in BCa tissues, urine and serum were consistent in our study. In conclusion, the sandwich ELISA successfully established in the present study was of high sensitivity and specificity in the detection of survivin expression. The results also indicated that survivin is a potential tumor marker for the surveillance of BCa and RCC.

摘要

Survivin,在大多数肿瘤中高度表达,但在大多数正常成人组织中不表达,已被确定具有重要的临床应用价值。在本研究中,我们使用 Survivin 特异性单克隆抗体(mAbs),旨在建立检测 Survivin 在膀胱癌(BCa)和肾细胞癌(RCC)患者的癌细胞系、血清样本、尿液样本和肿瘤组织中表达的方法,并评估 Survivin 作为肿瘤标志物在 BCa 和 RCC 监测中的疗效。首先,mAbs 被辣根过氧化物酶(HRP)标记,然后建立了 mAbs 和 HRP 标记的 mAbs 的夹心酶联免疫吸附试验(ELISA),以检测 BCa 和 RCC 患者血清和尿液样本中的 Survivin 表达,并用健康对照(HCs)样本进行比较。HRP 标记的 mAbs 也用于通过 Western blot 检测癌细胞系中的 Survivin 表达。还通过免疫组织化学评估了 BCa 患者肿瘤组织中的 Survivin 表达。结果表明,成功建立了夹心 ELISA,并且在尿液和血清样本中,与 HCs 相比,BCa 和 RCC 患者的 Survivin 表达明显更高(P<0.05),在尿液中更为明显。HRP-mAbs 可以识别癌细胞系中的 Survivin。Western blot 和免疫组化结果证实了 5637 BCa 细胞系以及 BCa 组织中 Survivin 的表达。此外,在本研究中,BCa 组织、尿液和血清中的 Survivin 表达一致。总之,本研究成功建立的夹心 ELISA 对 Survivin 表达的检测具有高灵敏度和特异性。结果还表明 Survivin 是 BCa 和 RCC 监测的潜在肿瘤标志物。

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