615 ADIPOPHILIN PROTEIN FOUND ONLY IN RCC PATIENT URINE: POTENTIAL USE FOR SCREENING AND CLASSIFICATION OF INCIDENTAL RENAL LESIONS (2013)

You have accessJournal of UrologyKidney Cancer: Basic Research (IV)1 Apr 2013615 ADIPOPHILIN PROTEIN FOUND ONLY IN RCC PATIENT URINE: POTENTIAL USE FOR SCREENING AND CLASSIFICATION OF INCIDENTAL RENAL LESIONS Shilpa Sreedharan, Rebecca S Arnold, Viraj A Master, Kenneth Ogan, David L Roberts, John G Pattaras, and John A Petros Shilpa SreedharanShilpa Sreedharan Atlanta, GA , Rebecca S ArnoldRebecca S Arnold Atlanta, GA , Viraj A MasterViraj A Master Atlanta, GA , Kenneth OganKenneth Ogan Atlanta, GA , David L RobertsDavid L Roberts Atlanta, GA , John G PattarasJohn G Pattaras Atlanta, GA , and John A PetrosJohn A Petros Atlanta, GA View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.166AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES There is no effective screening test for renal cell carcinoma (RCC). Two clinical scenarios could benefit from such a test: screening and evaluation of small renal lesions found incidentally during imaging. For screening, an accurate test could help in detecting RCC at a time when it is amenable to surgical cure. For the incidentally discovered small renal mass an accurate test could prevent needless surgery for benign lesions. A single report in the literature claims that there is no overlap between urine levels of adipose differentiation related protein (ADRP, or adipophilin) in patients with RCC and individuals without RCC (May Clin Proc 85:413, 2010). The objective of this project was to use a large archive of patient urine obtained at the time of nephrectomy for RCC in order to validate or refute this single report. METHODS Archived RCC patient urine and non-RCC negative control urine were centrifuged to remove debris and mixed with a protease inhibitor tablet before processing for Western blot analysis. Urinary creatinine concentrations were quantified by the Jaffe reaction (Nephron 16:31, 1976). Protein was precipitated from 100uL of fresh-spun urine in 1.5mL of ice-cold acetone-methanol (1:1) and dissolved in 1x SDS sample buffer for a final concentration of 2ug creatinine per uL. Cell lysates were prepared from human renal carcinoma cell lines Caki-1 and ACHN for positive controls. Blocked membranes were incubated with 1:200 dilution of anti-ADRP (H-80) antibody from Santa Cruz Biotechnology Inc, Santa Cruz, CA. Secondary antibody was HRP conjugated goat anti-rabbit (1:2000) and visualized by chemiluminescence. RESULTS Normal control urine failed to show any ADRP protein by western blot while both RCC patient urine protein and cell line lysates proteins were robustly positive for ADRP. There was no overlap between case (positive signal) and control (negative signal) making differentiation straightforward. CONCLUSIONS Our data confirms a previous report that the urine of patients with RCC contains easily detectable ADRP protein; even from archival specimens and that this is easily distinguished from control urine where there is absence of detectable ADRP. This clear distinction without overlap between case and control urine suggests that a clinically applicable diagnostic test for ADRP in the urine could be useful for both screening purposes and for the further diagnosis of incidentally discovered small renal lesions. Readily available cell lines appear to be appropriate positive controls for future assay development. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e251 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.Metrics Author Information Shilpa Sreedharan Atlanta, GA More articles by this author Rebecca S Arnold Atlanta, GA More articles by this author Viraj A Master Atlanta, GA More articles by this author Kenneth Ogan Atlanta, GA More articles by this author David L Roberts Atlanta, GA More articles by this author John G Pattaras Atlanta, GA More articles by this author John A Petros Atlanta, GA More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...