LAUSR

In vitro cancer research predominantly utilizes established cell lines of a chosen cancer type as phenotypic representations of various disease states, whether they be non-cancerous, non-invasive, highly metastatic, or derived from distinct cellular origins within an organ or tissue. While such methodology is useful to study cancer cells from diverse disease states, when one considers the tremendous complexity of intertumoral heterogeneity, it becomes problematic to use cell lines derived from distinct individuals with diverse malignancies to study the progression of tumorigenesis in a single cancer type. To address this need, a novel murine model was developed, composed of six cell lines isolated from progressively advanced tumors all derived from a single parental line (Ray et al., Exp. Cell Res. 340, 1-11). Each cell line should represent a snapshot of a particular stage of this cancer’s development, with the entire model providing a platform to study tumorigenic changes incurred as the transformed cells progressed from a localized tumor to an aggressively malignant growth. However, isolating primary cell lines can be an inefficient process, and a successfully isolated line may not accurately represent the dominant neoplastic cell type from a tumor. For that reason, it is necessary to characterize the cell lines from this model to confirm that the observed in vitro behavior of the cells matches the expected phenotype of the tissue from which it was derived. To this end, we performed an in vitro characterization of each cell line in the model. The three lines derived from distant metastatic tumors displayed significantly higher rates of two dimensional motility and invasive capacity as compared to the parental cell line, the line derived from a primary tumor or from a local metastasis. The distant metastatic lines also displayed elevated rates of extracellular acidification, an indication of increased rates of aerobic glycolysis. These measures demonstrated convincingly that the behavior of the cells in culture mirrored the expected phenotype of the tissue from which they were derived, thus validating the utility of the model. We then performed a comprehensive proteomic analysis via orbitrap tandem mass spectrometry, yielding expression data on over 3,000 proteins across all cell lines. This analysis yielded expected results that further validated the model, such as showing an increasing trend of expression of cancer-associated proteins like transcription factor AP-1 and decreasing trends of expression of tumor suppressors like protein NDRG1. However, it also revealed striking changes in protein expression in targets not currently implicated in tumorigenesis, thus providing novel targets for further examination. In total, this study validated a novel murine cell model as a valuable tool in the study of tumorigenesis, while providing a robust proteomic data set to guide future research into cancerous progression. Citation Format: Steven D. Scahill, Kelly Jean Sherman, Jessie Guidry, Harry J. Gould, Dennis Paul. In vitro characterization of a novel murine model of cancerous progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1684.