Abstract 3115: Physiologic comparisons of five bladder cancer cell lines
Authors: Richmond Bonnie L. , Damen Kyle , Moore Ryan , White Alex , Carter Julia H. 
Published: August 2018

Abstract
The use of tumor-derived cultured cells continues to be a major resource for understanding the biological activities of tumors and the effects of environmental factors and pharmacological agents on tumors. With comparison of data from the International Cancer Genome Consortium and the Cancer Genome Atlas comes the realization that some frequently used cells lines may not closely share genomic similarity with patient tumors, whereas less frequently used cell lines may share high similarity. For example, the bladder cancer cell line T24 (HTB-4) is more frequently used than other cell lines. We selected five commercially available bladder cancer cell lines, all derived from patient primary tumors, for a comparison of selected proteins, with the intent of determining how similar or dissimilar the cultured cells are. In selection of cultured cell lines, we considered reported grade of tumor from which the cell line was derived and the sex and age of the patient. We evaluated protein expression in newly confluent cultures and in sub-confluent cultures. The following cell lines were selected: RT4 (transitional cell papilloma), 5637 (grade II carcinoma), T24 (grade III carcinoma), HT 1376 (grade III carcinoma) and TCCSUP (grade IV carcinoma). We hypothesized that variations exist between bladder cancer cell lines isolated from low and high-grade carcinomas. Further, we identified physiological characteristics of the different bladder cancer cell lines at both newly confluent and sub confluent culture states as additional parameters for a bladder cancer model. Selection of an appropriate tissue culture cell line(s) and state of confluency are critical considerations in cellular research designed to understand the biological activities of tumors. Growth rates of the five cell lines were determined. Next, the basal-luminal status of the cell lines was determined using molecular subtyping markers (transcription factors GATA3 and p63, Cytokeratin 6, and Estrogen Receptor alpha). The presence of Cathepsin L, a cysteine protease, was evaluated. Further, we evaluated the expression of key proteins in the MAPK pathway (p38 MAPK, SAPK/JNK, and ERK1/2) as indicators of cellular stress. Last, the receptor status of Alpha 7 receptor, the putative receptor for nicotine, was determined. Our results indicate there is a difference in growth rates between the cell lines, with line 5637 having a delayed growth rate. Three cell lines were determined to be basal, and two cell lines were determined as mixed baso/luminal. The RT4 cell line had greater expression of cathepsin L. We observed no significant changes in the protein expression levels of the MAPK pathway nor of the alpha-7 receptor. Our results suggest that there are differences between the five bladder cancer cell lines suggesting that more than one cell line should be used and evaluated prior to beginning experiments to evaluate the potential effects of environmental factors and pharmacological agents.
Citation Format: Bonnie L. Richmond, Kyle Damen, Ryan Moore, Alex White, Julia H. Carter. Physiologic comparisons of five bladder cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3115.