title: Alternative CD44 splicing identifies epithelial prostate cancer cells from the mesenchymal counterparts.
abstract: An epithelial to mesenchymal transition (EMT) has been shown to be a necessary precursor to prostate cancer metastasis. Additionally, the differential expression and splicing of mRNAs has been identified as a key means to distinguish epithelial from mesenchymal cells by qPCR, western blotting and immunohistochemistry. However, few markers exist to differentiate between these cells by flow cytometry. We previously developed two cell lines, PC3-Epi (epithelial) and PC3-EMT (mesenchymal). RNAseq was used to determine the differential expression of membrane proteins on PC3-Epi/EMT. We used western blotting, qPCR and flow cytometry to validate the RNAseq results. CD44 was one of six membrane proteins found to be differentially spliced between epithelial and mesenchymal PC3 cells. Although total CD44 was positive in all PC3-Epi/EMT cells, PC3-Epi cells had a higher level of CD44v6 (CD44 variant exon 6). CD44v6 was able to differentiate epithelial from mesenchymal prostate cancer cells using either flow cytometry, western blotting or qPCR.
authors: Department of Urology, The James Buchanan Brady Urological Institute, Johns Hopkins University, Baltimore, MD, 21287, USA, jherna24@jhmi.edu.
authors: James R
authors: JR
authors: Hernandez
authors: John J
authors: JJ
authors: Kim
authors: James E
authors: JE
authors: Verdone
authors: Xin
authors: X
authors: Liu
authors: Gonzalo
authors: G
authors: Torga
authors: Kenneth J
authors: KJ
authors: Pienta
authors: Steven M
authors: SM
authors: Mooney
mesh_terms: D017398:Alternative Splicing / Q000235:genetics*; D045744:Cell Line, Tumor; D004847:Epithelial Cells / Q000473:pathology*; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics / Q000502:physiology; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics*; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000473:pathology*; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000473:pathology*
chemical_list: C497483:CD44 protein, human; D018960:Hyaluronan Receptors; D008565:Membrane Proteins