Abstract 3252: Establishment and characterization of a new squamous cell carcinoma cell line derived from human bladder cancer
Authors: Ozu Choichiro , Takeuchi Hisashi , Horiguchi Yutaka , Gondo Tatsuo , Namiki Kazunori , Yoshioka Kunihiko , Nakashima Jun , Hatano Tadashi , Tachibana Masaaki 
Published: February 2011

Abstract
Introduction: Squamous cell carcinoma (SCC) of the bladder is a rare malignancy that represents &lt;5% of bladder tumors diagnosed in new patients. The majority of the patients SCC presents with a poorly differentiated, muscle-invasive tumor with no previous episode of the urothelial carcinoma. Less than 10% of the patients present with distant metastasis. Even when the absence of distant metastases, the prognosis of patients with SCC of the bladder remains dismal because patients die of localized recurrence. The 5-year survival rate of the patients treated for SCC of the bladder was only 10.6%.
Therefore, the establishment of a bladder squamous cell carcinoma cell line is extremely important for SCC of the bladder cancer patients. As far as we are aware, however, no pure human SCC cell line of the bladder has ever been reported. This study describes our successful attempt to establish a cell line derived from inguinal lymph node metastasis from the bladder SCC.
Patient history: The patient was a 56-year-old Japanese woman. She underwent total cystectomy for bladder cancer in September 2007. The histopathological diagnosis was bladder squamous cell carcinoma (G2&gt;G3, T3bN0M0). She underwent adjuvant three courses of chemo therapy (M-VAC therapy) after surgery. However, local recurrence and multiple lymph node metastases were detected in January 2008. She underwent second operation as inguinal lymph node dissection.
Materials and Methods: Cells were cultured in RPMI medium 1640 supplemented with 10% fetal bovine serum, 50 U/ml penicillin and 50 μg/ml streptomycin. A portion of the inguinal tumor was studied histopathologically and the remainder was immersed in growth medium. The TMUU-08 cells transplanted into the subcutaneous of the back of five nude mice. Tumor cell suspension were applied on to slides using cytocentrifugation. FISH analysis was performed to detect the X- and Y- choromosome. The concentration of CEA, CA-125, CA-15-3 and SCC antigen in the conditioned medium with TMUU-08 cells were measured using enzyme immunoassay and electrochemiluminescence immunoassay. Subcutaneous tumors derived from TMUU-08 were used for immunohistochemical staining.
Results: Population doubling time of TMUU-08 was approximately 52 hrs. The subcutaneous tumors were histopathologically diagnosed as squamous cell carcinoma. The signals from subcutaneous tumor cells of Y, YY, X and XX were 0%, 0%, 43.8% and 55.6%, respectively. The concentration of SCC antigen in the conditioned medium was higher than the control. Immunohistochemical staining showed that more than 50% were positive against anti SCC antigen both in original tumor and in subcutaneous tumor section.
Conclusion: We have established and characterized new human bladder squamous cell carcinoma cell line. To our knowledge, the present study is the first establishment of pure human bladder squamous cell carcinoma cell line.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3252.