Identification of oxidized protein hydrolase as a potential prodrug target in prostate cancer

Abstract

BackgroundEsterases are often overexpressed in cancer cells and can have chiral specificities different from that of the corresponding normal tissues. For this reason, ester prodrugs could be a promising approach in chemotherapy. In this study, we focused on the identification and characterization of differentially expressed esterases between non-tumorigenic and tumorigenic prostate epithelial cells.MethodsCellular lysates from LNCaP, DU 145, and PC3 prostate cancer cell lines, tumorigenic RWPE-2 prostate epithelial cells, and non-tumorigenic RWPE-1 prostate epithelial cells were separated by native polyacrylamide gel electrophoresis (n-PAGE) and the esterase activity bands visualized using α-naphthyl acetate or α-naphthyl-N-acetylalaninate (ANAA) chiral esters and Fast Blue RR salt. The esterases were identified using nanospray LC/MS-MS tandem mass spectrometry and confirmed by Western blotting, native electroblotting, inhibition assays, and activity towards a known specific substrate. The serine protease/esterase oxidized protein hydrolase (OPH) was overexpressed in COS-7 cells to verify our results.ResultsThe major esterase observed with the ANAA substrates within the n-PAGE activity bands was identified as OPH. OPH (EC 3.4.19.1) is a serine protease/esterase and a member of the prolyl oligopeptidase family. We found that LNCaP lysates contained approximately 40% more OPH compared to RWPE-1 lysates. RWPE-2, DU145 and PC3 cell lysates had similar levels of OPH activity. OPH within all of the cell lysates tested had a chiral preference for the S-isomer of ANAA. LNCaP cells were stained more intensely with ANAA substrates than RWPE-1 cells and COS-7 cells overexpressing OPH were found to have a higher activity towards the ANAA and AcApNA than parent COS-7 cells.ConclusionsThese data suggest that prodrug derivatives of ANAA and AcApNA could have potential as chemotherapeutic agents for the treatment of prostate cancer tumors that overexpress OPH.

Combretastatin A4-camptothecin micelles as combination therapy for effective anticancer activity

“…The connection between the two anticancer drugs was through triazole group that are highly enzymatic and chemically stable. 38 Moreover, the direct linkage between the CA4, Cpt and linkers was through an ester bond that can be hydrolyzed easily by esterase enzyme that is usually overexpressed in cancer cells with a high intracellular concentration 39,40 in order to give a rapid release prole of the anticancer drugs.…”

Biological activity of pyrazole and imidazole-dehydroepiandrosterone derivatives on the activity of 17β-hydroxysteroid dehydrogenase

“…With the exception of 4e, these steroids also exhibited pharmacological activity. The overall results suggested compound 4e was not hydrolyzed by the specific esterase present in the membrane fraction of prostate homogenates 27 . This esterase has been previously identified as an oxidized protein hydrolase (OPH), which is overexpressed in the tumorigenic LNCaP cell line.…”

“…This esterase has been previously identified as an oxidized protein hydrolase (OPH), which is overexpressed in the tumorigenic LNCaP cell line. OPH is a serine esterase/protease that has a well-characterized esterase activity towards a-naphthyl butyrate and an exopeptidase activity for removing the N-terminally acetylated amino acid residues from peptides/proteins 27 . Figure 5.…”

microRNA-99a inhibits cell proliferation, colony formation ability, migration and invasion by targeting fibroblast growth factor receptor 3 in prostate cancer

“…Prostate cancer remains one of the most prevalent types of cancer in males (1). In the USA and Europe, prostate cancer is currently the second most common cause of cancer mortality in males >40 years of age, and the third most common cause of cancer-associated mortality in males (2,3). A total of 238,590 novel cases and 29,720 mortalities due to prostate cancer were estimated for 2013 in the USA (2).…”

“…In the USA and Europe, prostate cancer is currently the second most common cause of cancer mortality in males >40 years of age, and the third most common cause of cancer-associated mortality in males (2,3). A total of 238,590 novel cases and 29,720 mortalities due to prostate cancer were estimated for 2013 in the USA (2). Although the etiology of this malignancy remains to be elucidated, several risk factors have been identified that are considered to contribute to prostate carcinogenesis, including hereditary and environmental components; however, age, race and family history are the only well-established risk factors (4).…”