Investigation of PKHD1L1's Localization and Potential Function as a Tumor Suppressor

Abstract

The Polycystic Kidney and Hepatic Disease 1‐like 1 (PKHD1L1) gene located on human chromosome 8 has been shown to have a high frequency of mutation in multiple types of epithelial cancers, such as skin, lung, and bladder cancer, according to data generated by the TCGA Research Network; its mutation rate in bladder urothelial carcinoma (11.41%) was ranked 65 among 19,578 genes analyzed. Furthermore, the TCGA data demonstrated that PKHD1L1 gene expression was diminished compared to normal tissues in those epithelial tumors. Our approach was to examine PKHD1L1's potential tumor suppression functions in bladder cancer cell line UM‐UC‐3. We engineered a CRISPR‐Cas9 PKHD1L1‐gRNA expression vector, which was then transfected into UM‐UC‐3 cells. After selection and establishment of clonal cell lines, mutation of full‐length PKHD1L1 was confirmed to be successful via TIDE analysis showing complete, frame‐shift disruptions. We performed wound healing assays, MTS assays, and colony formation assays to examine the impact of full‐length PKHD1L1 deficiency on cell motility and proliferative capacity. Wound healing assays and MTS assays showed no significant difference due to PKHD1L1 deficiency. Preliminary studies suggest PKHD1L1 deficiency may result in larger and more numerous colonies in the colony formation assay. We are currently investigating the subcellular localization of PKHD1L1 protein through immunocytochemistry. With further investigation, our research can provide significant insight into the role of PKHD1L1 in cancer development and progression.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.