Presentation: 2025 ND EPSCoR Annual conference 

October 21, 2025, NDSU Memorial Union, Fargo, North Dakota

Analysis of SPARC, CDH1, and CDH2 Expression in in vitro Bladder Cancer Media Swap Models

Bladder cancer is the 10th most common type of cancer in the world. In 2024, there were 83,190 new cases in the United States alone, a number that has shown steady increase over the years. Bladder cancer is strongly linked to environmental toxin exposure, such as tobacco smoking or heavy metals like cadmium, which can be found in soil and water. SPARC, a matricellular protein, and E-Cadherin (CDH1), and N-Cadherin, (CDH2), cell adhesion proteins, are all important in bladder cancer. Expression of SPARC in bladder cancer has been shown to be reduced in low grades and following cadmium exposure, promoting a cancerous phenotype. Both CDH1 and CDH2 are hallmarks of epithelial-mesenchymal transition, with a cadherin switch between CDH1 and CDH2, promoting cancer progression. The purpose of our project was to examine if expression of these genes when swapping the culture media to optimize co-culture conditions. Non-tumorigenic UROtsa and tumorigenic Cd2+, RT4, and T24 cell lines were cultured along with CAF cells, in their respective media formulations. These cells were then cultured in a media swap set up from 0% to 100% by increments of 25%. All wells were harvested, RNA was extracted, synthesized to cDNA, and used for RT-PCR. Results showed that the expression of all three genes stayed consistent throughout media swap increments, retaining their profile despite the new media. Results also showed expected expression profile across in vitro models, further supporting optimized conditions. We are currently repeating our bladder cell/CAF co-culture set up and optimizing 3D culture conditions.