Proteotranscriptomic Measurements of E6-Associated Protein (E6AP) Targets in DU145 Prostate Cancer Cells

Twishi Gulati, Cheng Huang, Franco Caramia, Dinesh Raghu, Piotr J. Paul, Robert J.A. Goode, Simon P. Keam, Scott G. Williams, Sue Moody Haupt, Oded Kleifeld, Ralf B. Schittenhelm, Cristina Gamell, Ygal Haupt

Research output: Contribution to journalArticleResearchpeer-review

1 Citation (Scopus)

Abstract

Prostate cancer is a common cause of cancer-related death in men. E6AP (E6-Associated Protein), an E3 ubiquitin ligase and a transcription cofactor, is elevated in a subset of prostate cancer patients. Genetic manipulations of E6AP in prostate cancer cells expose a role of E6AP in promoting growth and survival of prostate cancer cells in vitro and in vivo. However, the effect of E6AP on prostate cancer cells is broad and it cannot be explained fully by previously identified tumor suppressor targets of E6AP, promyelocytic leukemia protein and p27. To explore additional players that are regulated downstream of E6AP, we combined a transcriptomic and proteomic approach. We identified and quantified 16,130 transcripts and 7,209 proteins in castration resistant prostate cancer cell line, DU145. A total of 2,763 transcripts and 308 proteins were significantly altered on knockdown of E6AP. Pathway analyses supported the known phenotypic effects of E6AP knockdown in prostate cancer cells and in parallel exposed novel potential links of E6AP with cancer metabolism, DNA damage repair and immune response. Changes in expression of the top candidates were confirmed using real-time polymerase chain reaction. Of these, clusterin, a stress-induced chaperone protein, commonly deregulated in prostate cancer, was pursued further. Knockdown of E6AP resulted in increased clusterin transcript and protein levels in vitro and in vivo. Concomitant knockdown of E6AP and clusterin supported the contribution of clusterin to the phenotype induced by E6AP. Overall, results from this study provide insight into the potential biological pathways controlled by E6AP in prostate cancer cells and identifies clusterin as a novel target of E6AP.

Original languageEnglish
Pages (from-to)1170-1183
Number of pages14
JournalMolecular & Cellular Proteomics
Volume17
Issue number6
DOIs
Publication statusPublished - 1 Jun 2018

Cite this

Gulati, Twishi ; Huang, Cheng ; Caramia, Franco ; Raghu, Dinesh ; Paul, Piotr J. ; Goode, Robert J.A. ; Keam, Simon P. ; Williams, Scott G. ; Haupt, Sue Moody ; Kleifeld, Oded ; Schittenhelm, Ralf B. ; Gamell, Cristina ; Haupt, Ygal. / Proteotranscriptomic Measurements of E6-Associated Protein (E6AP) Targets in DU145 Prostate Cancer Cells. In: Molecular & Cellular Proteomics. 2018 ; Vol. 17, No. 6. pp. 1170-1183.
@article{af7e628ddfd94ff4b9c5c848dfb68aa7,
title = "Proteotranscriptomic Measurements of E6-Associated Protein (E6AP) Targets in DU145 Prostate Cancer Cells",
abstract = "Prostate cancer is a common cause of cancer-related death in men. E6AP (E6-Associated Protein), an E3 ubiquitin ligase and a transcription cofactor, is elevated in a subset of prostate cancer patients. Genetic manipulations of E6AP in prostate cancer cells expose a role of E6AP in promoting growth and survival of prostate cancer cells in vitro and in vivo. However, the effect of E6AP on prostate cancer cells is broad and it cannot be explained fully by previously identified tumor suppressor targets of E6AP, promyelocytic leukemia protein and p27. To explore additional players that are regulated downstream of E6AP, we combined a transcriptomic and proteomic approach. We identified and quantified 16,130 transcripts and 7,209 proteins in castration resistant prostate cancer cell line, DU145. A total of 2,763 transcripts and 308 proteins were significantly altered on knockdown of E6AP. Pathway analyses supported the known phenotypic effects of E6AP knockdown in prostate cancer cells and in parallel exposed novel potential links of E6AP with cancer metabolism, DNA damage repair and immune response. Changes in expression of the top candidates were confirmed using real-time polymerase chain reaction. Of these, clusterin, a stress-induced chaperone protein, commonly deregulated in prostate cancer, was pursued further. Knockdown of E6AP resulted in increased clusterin transcript and protein levels in vitro and in vivo. Concomitant knockdown of E6AP and clusterin supported the contribution of clusterin to the phenotype induced by E6AP. Overall, results from this study provide insight into the potential biological pathways controlled by E6AP in prostate cancer cells and identifies clusterin as a novel target of E6AP.",
author = "Twishi Gulati and Cheng Huang and Franco Caramia and Dinesh Raghu and Paul, {Piotr J.} and Goode, {Robert J.A.} and Keam, {Simon P.} and Williams, {Scott G.} and Haupt, {Sue Moody} and Oded Kleifeld and Schittenhelm, {Ralf B.} and Cristina Gamell and Ygal Haupt",
year = "2018",
month = "6",
day = "1",
doi = "10.1074/mcp.RA117.000504",
language = "English",
volume = "17",
pages = "1170--1183",
journal = "Molecular & Cellular Proteomics",
issn = "1535-9476",
publisher = "American Society for Biochemistry and Molecular Biology",
number = "6",

}

Gulati, T, Huang, C, Caramia, F, Raghu, D, Paul, PJ, Goode, RJA, Keam, SP, Williams, SG, Haupt, SM, Kleifeld, O, Schittenhelm, RB, Gamell, C & Haupt, Y 2018, 'Proteotranscriptomic Measurements of E6-Associated Protein (E6AP) Targets in DU145 Prostate Cancer Cells', Molecular & Cellular Proteomics, vol. 17, no. 6, pp. 1170-1183. https://doi.org/10.1074/mcp.RA117.000504

Proteotranscriptomic Measurements of E6-Associated Protein (E6AP) Targets in DU145 Prostate Cancer Cells. / Gulati, Twishi; Huang, Cheng; Caramia, Franco; Raghu, Dinesh; Paul, Piotr J.; Goode, Robert J.A.; Keam, Simon P.; Williams, Scott G.; Haupt, Sue Moody; Kleifeld, Oded; Schittenhelm, Ralf B.; Gamell, Cristina; Haupt, Ygal.

In: Molecular & Cellular Proteomics, Vol. 17, No. 6, 01.06.2018, p. 1170-1183.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Proteotranscriptomic Measurements of E6-Associated Protein (E6AP) Targets in DU145 Prostate Cancer Cells

AU - Gulati, Twishi

AU - Huang, Cheng

AU - Caramia, Franco

AU - Raghu, Dinesh

AU - Paul, Piotr J.

AU - Goode, Robert J.A.

AU - Keam, Simon P.

AU - Williams, Scott G.

AU - Haupt, Sue Moody

AU - Kleifeld, Oded

AU - Schittenhelm, Ralf B.

AU - Gamell, Cristina

AU - Haupt, Ygal

PY - 2018/6/1

Y1 - 2018/6/1

N2 - Prostate cancer is a common cause of cancer-related death in men. E6AP (E6-Associated Protein), an E3 ubiquitin ligase and a transcription cofactor, is elevated in a subset of prostate cancer patients. Genetic manipulations of E6AP in prostate cancer cells expose a role of E6AP in promoting growth and survival of prostate cancer cells in vitro and in vivo. However, the effect of E6AP on prostate cancer cells is broad and it cannot be explained fully by previously identified tumor suppressor targets of E6AP, promyelocytic leukemia protein and p27. To explore additional players that are regulated downstream of E6AP, we combined a transcriptomic and proteomic approach. We identified and quantified 16,130 transcripts and 7,209 proteins in castration resistant prostate cancer cell line, DU145. A total of 2,763 transcripts and 308 proteins were significantly altered on knockdown of E6AP. Pathway analyses supported the known phenotypic effects of E6AP knockdown in prostate cancer cells and in parallel exposed novel potential links of E6AP with cancer metabolism, DNA damage repair and immune response. Changes in expression of the top candidates were confirmed using real-time polymerase chain reaction. Of these, clusterin, a stress-induced chaperone protein, commonly deregulated in prostate cancer, was pursued further. Knockdown of E6AP resulted in increased clusterin transcript and protein levels in vitro and in vivo. Concomitant knockdown of E6AP and clusterin supported the contribution of clusterin to the phenotype induced by E6AP. Overall, results from this study provide insight into the potential biological pathways controlled by E6AP in prostate cancer cells and identifies clusterin as a novel target of E6AP.

AB - Prostate cancer is a common cause of cancer-related death in men. E6AP (E6-Associated Protein), an E3 ubiquitin ligase and a transcription cofactor, is elevated in a subset of prostate cancer patients. Genetic manipulations of E6AP in prostate cancer cells expose a role of E6AP in promoting growth and survival of prostate cancer cells in vitro and in vivo. However, the effect of E6AP on prostate cancer cells is broad and it cannot be explained fully by previously identified tumor suppressor targets of E6AP, promyelocytic leukemia protein and p27. To explore additional players that are regulated downstream of E6AP, we combined a transcriptomic and proteomic approach. We identified and quantified 16,130 transcripts and 7,209 proteins in castration resistant prostate cancer cell line, DU145. A total of 2,763 transcripts and 308 proteins were significantly altered on knockdown of E6AP. Pathway analyses supported the known phenotypic effects of E6AP knockdown in prostate cancer cells and in parallel exposed novel potential links of E6AP with cancer metabolism, DNA damage repair and immune response. Changes in expression of the top candidates were confirmed using real-time polymerase chain reaction. Of these, clusterin, a stress-induced chaperone protein, commonly deregulated in prostate cancer, was pursued further. Knockdown of E6AP resulted in increased clusterin transcript and protein levels in vitro and in vivo. Concomitant knockdown of E6AP and clusterin supported the contribution of clusterin to the phenotype induced by E6AP. Overall, results from this study provide insight into the potential biological pathways controlled by E6AP in prostate cancer cells and identifies clusterin as a novel target of E6AP.

UR - http://www.scopus.com/inward/record.url?scp=85048183514&partnerID=8YFLogxK

U2 - 10.1074/mcp.RA117.000504

DO - 10.1074/mcp.RA117.000504

M3 - Article

VL - 17

SP - 1170

EP - 1183

JO - Molecular & Cellular Proteomics

JF - Molecular & Cellular Proteomics

SN - 1535-9476

IS - 6

ER -