Gills Jessie, Moret Ravan, Zhang Xin, Nelson John, Maresh Grace, Hellmers Linh, Canter Daniel, Hudson M'Liss, Halat Shams, Matrana Marc, Marino Michael P, Reiser Jakob, Shuh Maureen, Laborde Eric, Latsis Maria, Talwar Sunil, Bardot Stephen, Li Li
Department of Urology, Ochsner Clinic Foundation, New Orleans, LA, USA.
Institution of Translational Research, Ochsner Clinic Foundation, New Orleans, LA, USA.
Oncotarget. 2018 Aug 24;9(66):32718-32729. doi: 10.18632/oncotarget.26024.
High-grade urothelial cell carcinoma of the bladder has a poor prognosis when lymph nodes are involved. Despite curative therapy for clinically-localized disease, over half of the muscle-invasive urothelial cell carcinoma patients will develop metastases and die within 5 years. There are currently no described xenograft models that consistently mimic urothelial cell carcinoma metastasis. To develop a patient-derived orthotopic xenograft model to mimic clinical urothelial cell carcinoma progression to metastatic disease, the urothelial cell carcinoma cell line UM-UC-3 and two urothelial cell carcinoma patient specimens were doubly tagged with Luciferase/RFP and were intra-vesically (IB) instilled into NOD/SCID mice with or without lymph node stromal cells (HK cells). Mice were monitored weekly with bioluminescence imaging to assess tumor growth and metastasis. Primary tumors and organs were harvested for bioluminescence imaging, weight, and formalin-fixed for hematoxylin and eosin and immunohistochemistry staining. In this patient-derived orthotopic xenograft model, xenograft tumors showed better implantation rates than currently reported using other models. Xenograft tumors histologically resembled pre-implanted primary specimens from patients, presenting muscle-invasive growth patterns. In the presence of HK cells, tumor formation, tumor angiogenesis, and distant organ metastasis were significantly enhanced in both UM-UC-3 cells and patient-derived specimens. Thus, we established a unique, reproducible patient-derived orthotopic xenograft model using human high-grade urothelial cell carcinoma cells and lymph node stromal cells. It allows for investigating the mechanism involved in tumor formation and metastasis, and therefore it is useful for future testing the optimal sequence of conventional drugs or the efficacy of novel therapeutic drugs.
膀胱高级别尿路上皮细胞癌在出现淋巴结转移时预后较差。尽管对临床局限性疾病进行了根治性治疗，但超过一半的肌层浸润性尿路上皮细胞癌患者仍会发生转移并在5年内死亡。目前尚无能够持续模拟尿路上皮细胞癌转移的异种移植模型。为了建立一种源自患者的原位异种移植模型以模拟临床尿路上皮细胞癌向转移性疾病的进展，将尿路上皮细胞癌细胞系UM-UC-3和两份尿路上皮细胞癌患者标本用荧光素酶/RFP进行双重标记，并通过膀胱内（IB）注射到有或没有淋巴结基质细胞（HK细胞）的NOD/SCID小鼠体内。每周用生物发光成像监测小鼠，以评估肿瘤生长和转移情况。采集原发性肿瘤和器官进行生物发光成像、称重，并进行福尔马林固定以进行苏木精和伊红染色以及免疫组织化学染色。在这个源自患者的原位异种移植模型中，异种移植肿瘤的植入率比目前使用其他模型报道的更好。异种移植肿瘤在组织学上类似于患者植入前的原发性标本，呈现肌层浸润性生长模式。在存在HK细胞的情况下，UM-UC-3细胞和源自患者的标本中的肿瘤形成、肿瘤血管生成和远处器官转移均显著增强。因此，我们使用人高级别尿路上皮细胞癌细胞和淋巴结基质细胞建立了一种独特的、可重复的源自患者的原位异种移植模型。它有助于研究肿瘤形成和转移所涉及的机制，因此对于未来测试传统药物的最佳给药顺序或新型治疗药物的疗效很有用。
