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• W2004104079 abstract "You have accessJournal of UrologyBladder Cancer: Basic Research1 Apr 20111070 DEVELOPMENT OF A RAPAMYCIN SENSITIVE CELL LINE FROM UROTHELIAL CARCINOMA WITH SARCOMATOID FEATURES Edward Diaz, Christina Ching, and Donna Hansel Edward DiazEdward Diaz Cleveland, OH More articles by this author , Christina ChingChristina Ching Cleveland, OH More articles by this author , and Donna HanselDonna Hansel Cleveland, OH More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2011.02.1107AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Multiple commercial cell lines for urothelial carcinoma (UC) exist, but few were developed from aggressive variants of urothelial carcinoma, such as those with sarcomatoid or squamous features. There are also very few urothelial cell lines that can be sustained in a serum free environment. Recent studies support a relationship between regulation of the mammalian target of rapamycin (mTOR) pathway and carcinogenesis in UC. Our objective was to develop a serum independent cell line from an aggressive variant of urothelial carcinoma, and assess the effect of mTOR inhibition on these cells. METHODS The primary specimen was obtained from a 67 yo caucasian female with high grade UC with sarcomatoid features and squamous differentiation. Cells were immortalized using two rounds of cell propagation in an immune deficient mouse model. The final tumor was harvested from the mouse, dissociated, and grown in serum free conditions supplemented with growth factors. The cells were then tested for expression of cell markers found in aggressive UC (CK7, CK20, p63, p53, and Ker903) using a cell block protocol and immunostaining. Sensitivity of the cell line to mTOR inhibition was then tested in vivo using a double arm trial of placebo vs rapamycin in mice with established tumors. RESULTS Cells grew in a monolayer adherent to standard tissue culture plates. Morphology and viability was maintained after multiples passages within serum free conditions (Over 20 passages). Cell block revealed approximately 30% staining for p63, 100% staining for CK7, 0% staining for CK20, 100% staining for p53, and 40% staining for Ker903, confirming the cell line's human urothelial origin. The cell line demonstrated tumorigenicity in the mouse xenograft model. Single injection of 3 million cells resulted in the growth of a tumor with max 1 cm dimension after 31 days from inoculation. Tumors derived from this cell line demonstrated in vivo sensitivity to the mTOR inhibitor, rapamycin. Starting average tumor volume was: 231.5 cm3 for rapamycin arm 302.4 cm3 for placebo arm. Average tumor volume after treatment was: 336 cm3 for rapamycin arm, and 682 cm3 for placebo arm. No metastatic disease was present in either treatment group. CONCLUSIONS This provides a unique urothelial cancer cell line derived from an aggressive variant of UC and one that can be sustained in serum free conditions. Preliminary results in this study suggest mTOR signaling is important to the proliferative potential of this cell line, and possibly to this aggressive variant of UC. © 2011 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 185Issue 4SApril 2011Page: e430 Advertisement Copyright & Permissions© 2011 by American Urological Association Education and Research, Inc.MetricsAuthor Information Edward Diaz Cleveland, OH More articles by this author Christina Ching Cleveland, OH More articles by this author Donna Hansel Cleveland, OH More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ..." @default.
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• W2004104079 title "1070 DEVELOPMENT OF A RAPAMYCIN SENSITIVE CELL LINE FROM UROTHELIAL CARCINOMA WITH SARCOMATOID FEATURES" @default.
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