TY - JOUR
T1 - p66Shc regulates migration of castration-resistant prostate cancer cells
AU - Ingersoll, Matthew A.
AU - Chou, Yu Wei
AU - Lin, Jamie S.
AU - Yuan, Ta Chun
AU - Miller, Dannah R.
AU - Xie, Yan
AU - Tu, Yaping
AU - Oberley-Deegan, Rebecca E.
AU - Batra, Surinder K.
AU - Lin, Ming Fong
N1 - Funding Information:
This work was supported in part by the National Institute of Health [R01 CA88184, UO1 CA185148], Department of Defense [PC121645 and PC141559], the University of Nebraska Medical Center Bridge Fund, University of Nebraska Food for Health Collaboration Initiative Seed Grant from Nebraska EPSCoR, and UNMC Graduate Studies Fellowship, the Purdue Pharma Scholars Award, Epply Cancer Biology Training Program [T32CA009476], the Ministry of Science and Technology, Taiwan [MOST 106-2320-B-182A-001] and Kaohsiung Chang Gung Memorial Hospital, Taiwan [CMRPG8G0341] and Ben J. Lipps Research Fellowship Award from the ASN Foundation for Kidney Research. Support for the UNMC Advanced Microscopy Core Facility was provided by the Nebraska Research Initiative, the Fred and Pamela Buffett Cancer Center Support Grant [P30CA036727], and an Institutional Development Award (IDeA) from the NIGMS of the NIH [P30GM106397].
Funding Information:
This work was supported in part by the National Institute of Health [ R01 CA88184 , UO1 CA185148 ], Department of Defense [ PC121645 and PC141559 ], the University of Nebraska Medical Center Bridge Fund, University of Nebraska Food for Health Collaboration Initiative Seed Grant from Nebraska EPSCoR, and UNMC Graduate Studies Fellowship, the Purdue Pharma Scholars Award, Epply Cancer Biology Training Program [ T32CA009476 ], the Ministry of Science and Technology, Taiwan [ MOST 106-2320-B-182A-001 ] and Kaohsiung Chang Gung Memorial Hospital, Taiwan [ CMRPG8G0341 ] and Ben J. Lipps Research Fellowship Award from the ASN Foundation for Kidney Research. Support for the UNMC Advanced Microscopy Core Facility was provided by the Nebraska Research Initiative, the Fred and Pamela Buffett Cancer Center Support Grant [P30CA036727], and an Institutional Development Award (IDeA) from the NIGMS of the NIH [P30GM106397].
Publisher Copyright:
© 2018 Elsevier Inc.
Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.
PY - 2018/6
Y1 - 2018/6
N2 - Metastatic castration-resistant (CR) prostate cancer (PCa) is a lethal disease for which no effective treatment is currently available. p66Shc is an oxidase previously shown to promote androgen-independent cell growth through generation of reactive oxygen species (ROS) and is elevated in clinical PCa and multiple CR PCa cell lines. We hypothesize p66Shc also increases the migratory activity of PCa cells through ROS and investigate the associated mechanism. Using the transwell assay, our study reveals that the level of p66Shc protein correlates with cell migratory ability across several PCa cell lines. Furthermore, we show hydrogen peroxide treatment induces migration of PCa cells that express low levels of p66Shc in a dose-dependent manner, while antioxidants inhibit migration. Conversely, PCa cells that express high levels of endogenous p66Shc or by cDNA transfection possess increased cell migration which is mitigated upon p66Shc shRNA transfection or expression of oxidase-deficient dominant-negative p66Shc W134F mutant. Protein microarray and immunoblot analyses reveal multiple proteins, including ErbB-2, AKT, mTOR, ERK, FOXM1, PYK2 and Rac1, are activated in p66Shc-elevated cells. Their involvement in PCa migration was examined using respective small-molecule inhibitors. The role of Rac1 was further validated using cDNA transfection and, significantly, p66Shc is found to promote lamellipodia formation through Rac1 activation. In summary, the results of our current studies clearly indicate p66Shc also regulates PCa cell migration through ROS-mediated activation of migration-associated proteins, notably Rac1.
AB - Metastatic castration-resistant (CR) prostate cancer (PCa) is a lethal disease for which no effective treatment is currently available. p66Shc is an oxidase previously shown to promote androgen-independent cell growth through generation of reactive oxygen species (ROS) and is elevated in clinical PCa and multiple CR PCa cell lines. We hypothesize p66Shc also increases the migratory activity of PCa cells through ROS and investigate the associated mechanism. Using the transwell assay, our study reveals that the level of p66Shc protein correlates with cell migratory ability across several PCa cell lines. Furthermore, we show hydrogen peroxide treatment induces migration of PCa cells that express low levels of p66Shc in a dose-dependent manner, while antioxidants inhibit migration. Conversely, PCa cells that express high levels of endogenous p66Shc or by cDNA transfection possess increased cell migration which is mitigated upon p66Shc shRNA transfection or expression of oxidase-deficient dominant-negative p66Shc W134F mutant. Protein microarray and immunoblot analyses reveal multiple proteins, including ErbB-2, AKT, mTOR, ERK, FOXM1, PYK2 and Rac1, are activated in p66Shc-elevated cells. Their involvement in PCa migration was examined using respective small-molecule inhibitors. The role of Rac1 was further validated using cDNA transfection and, significantly, p66Shc is found to promote lamellipodia formation through Rac1 activation. In summary, the results of our current studies clearly indicate p66Shc also regulates PCa cell migration through ROS-mediated activation of migration-associated proteins, notably Rac1.
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U2 - 10.1016/j.cellsig.2018.02.008
DO - 10.1016/j.cellsig.2018.02.008
M3 - Article
C2 - 29462661
AN - SCOPUS:85042387227
VL - 46
SP - 1
EP - 14
JO - Cellular Signalling
JF - Cellular Signalling
SN - 0898-6568
ER -