Du Yiqing, Wang Lei, Wang Weiyi, Guo Ting, Zhang Mengzhao, Zhang Pu, Zhang Yanning, Wu Kaijie, Li Axing, Wang Xinyang, He Jiankang, Fan Jinhai
J Biomed Nanotechnol. 2018 Jan 1;14(1):161-167. doi: 10.1166/jbn.2018.2499.
As a novel cell-permeable peptide, polyarginine (R11) showed great potential as contrast agent to target bladder cancer (BCa) for therapeutic applications. However, its diagnostic ability and uptake efficiency between BCa and normal bladder tissues is unknown. In this study, we investigated the feasibility of R11 conjugated with fluorescein isothiocyanate (FITC-R11) for detecting BCa in clinical practice.
FITC-R11 was synthesized and incubated with BCa cell lines (T24, 5637, RT4), normal immortalized human bladder epithelial cell line (SVHUC) and clinical specimens from BCa and benign prostate hyperplasia patients. The uptake efficiency was determined by the mean values of relative FITC intensity. Furthermore, FITC-R11 was intravesically injected into the athymic nude mice bearing orthotopic T24-t tumors and pulmonary metastasis of bladder tumor mice models, the fluorescence intensity of bladder tumors and normal bladder tissues was examined, as well as lung tissues.
After incubation with FITC-R11, the fluorescence intensity of T24, 5637, RT4 and SVHUC cell line was 64678.56 ± 9699.27, 46456.22 ± 2588.06, 33802.02 ± 429.72 and 17785.01 ± 1704.36, respectively (P < 0.05). In the athymic nude mice bearing orthotopic T24-t tumors, FITC-R11 showed elevated accumulation in the bladder tumors compared with normal bladder tissues, FITC-R11 was also accumulated in the lung of pulmonary metastasis mice. Moreover, the uptake efficiency of FITC-R11 in patients' BCa tissues was much higher than in normal bladder tissues (6441.95 vs. 1196.92, P < 0.05), as well as the urine samples of BCa patients and benign prostate hyperplasia patients (30250.37 vs. 4948.42, P < 0.05).
As compared with normal bladder tissue, FITC-R11 is a more specific molecular probe for BCa, and has the potential application in clinical practice.
作为一种新型的细胞穿透肽，聚精氨酸（R11）作为造影剂在靶向膀胱癌（BCa）治疗应用方面显示出巨大潜力。然而，其在BCa与正常膀胱组织之间的诊断能力和摄取效率尚不清楚。在本研究中，我们探讨了异硫氰酸荧光素偶联的R11（FITC-R11）在临床实践中检测BCa的可行性。
合成FITC-R11，并将其与BCa细胞系（T24、5637、RT4）、永生化人正常膀胱上皮细胞系（SVHUC）以及来自BCa患者和良性前列腺增生患者的临床标本共同孵育。通过相对FITC强度的平均值来确定摄取效率。此外，将FITC-R11经膀胱内注射到原位接种T24-t肿瘤的无胸腺裸鼠和膀胱肿瘤肺转移小鼠模型中，检测膀胱肿瘤、正常膀胱组织以及肺组织的荧光强度。
与FITC-R11孵育后，T24、5637、RT4和SVHUC细胞系的荧光强度分别为64678.56±9699.27、46456.22±2,588.06、33802.02±429.72和17785.01±1704.36（P<0.05）。在原位接种T24-t肿瘤的无胸腺裸鼠中，与正常膀胱组织相比，FITC-R11在膀胱肿瘤中的蓄积增加，FITC-R11也在肺转移小鼠的肺中蓄积。此外，FITC-R11在患者BCa组织中的摄取效率远高于正常膀胱组织（6441.95对1196.92，P<0.05），在BCa患者和良性前列腺增生患者的尿液样本中也是如此（30250.37对4948.42，P<0.05）。
与正常膀胱组织相比，FITC-R11是一种针对BCa的更具特异性的分子探针，具有临床实践中的潜在应用价值。
