Investigating the Expression of Wnt5a Isoforms in Urothelial Carcinoma

Abstract

It is projected that there will be over 81,000 new bladder cancer diagnoses in 2018 in the United States alone. In fact, 1 in 27 men will develop bladder cancer in their lifetime. Despite its prevalence, little is known about this cancer's molecular mechanism. Research performed both at Ohio University and by other scientists has linked a protein, Wnt5a, to urothelial carcinoma (UC), which is the most common form of bladder cancer. This protein has been shown to be involved in an array of diseases, including several types of cancer. Recent research has suggested that two Wnt5a isoforms, Wnt5a‐Short (S) and Wnt5a‐Long (L), may play differing roles in a variety of cancers, with Wnt5a‐S promoting cellular proliferation and Wnt5a‐L suppressing cellular proliferation. Based on this and the research linking Wnt5a to UC, we want to investigate the roles of the Wnt5a isoforms in UC cell lines and human UC biopsies. We extracted RNA from human UC cell lines derived from low and high grade tumors to quantify the amount of each Wnt5a isoform and total Wnt5a expression using real‐time reverse transcription / polymerase chain reaction (RT‐RT/PCR). Trends in expression for Wnt5a‐L and Wnt5a‐Total were extremely similar, suggesting that Wnt5a‐L is the predominantly expressed isoform in urothelial cells. Wnt5a‐L RNA expression was highest in a cell line derived from a high grade tumor but also lowest in a cell line derived from a second high grade tumor. In contrast, expression of Wnt5a‐S RNA was highest in cell lines derived from high grade tumors in comparison to low grade tumors. Linking this to previous research published by our lab, the pattern of Wnt5a‐S expression was similar to that of vimentin, a marker of epithelial mesenchymal transition (EMT) linked to metastasis. We are currently performing immunohistochemistry (IHC) and laser microdissection (LMD) to visualize the location of Wnt5a isoform expression in human UC biopsies from both low grade and high grade tumors. Our hypothesis is that high grade tumors will express higher levels of the Wnt5a‐S isoform than low grade tumors; in contrast, high grade tumors will express lower levels of Wnt5a‐L than low grade tumors. An affirmative outcome of our hypothesis would suggest that the Wnt5a‐S isoform promotes oncogenic properties in UC, while the Wnt5a‐L isoform promotes tumor suppressive properties. Future experiments will be aimed at determining the molecular pathway(s) through which each isoform acts in order to advance diagnostic, prognostic, and therapeutic approaches to UC.Support or Funding InformationFunding and support were provided in part by Ohio University Heritage College of Osteopathic Medicine and its Research and Scholarly Advancement Fellowship (RSAF), the Diabetes Institute at Ohio University, as well as by an Ohio University Student Enhancement Award (SEA).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.