Itch, an E3 ubiquitin ligase, influences downstream receptor tyrosine kinase signalling in the colorectal cancer cell line, HCT116

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Receptor tyrosine kinase signalling is an important driver of cancer cell development. Deregulation of the receptor tyrosine kinases EGFR and Met is associated with cancer formation and progression to metastasis. Recently, mutations in Met that affect endocytic trafficking have been shown to promote cell migration, metastasis and tumorigenesis due to increased endosomal signalling. Thus, Met endocytic trafficking regulates signalling pathways; such as ERK1/2, STAT3, Gab1 and Rac1; that are implicated in cell survival, invasion and metastasis. Advancements in our understanding of regulation of EGFR and Met signalling is essential in order to understand their role in cancer progression. Ubiquitination regulates both receptor degradation and membrane trafficking. Here we investigated the role of the ubiquitin E3 ligase Itch in endocytosis and EGFR and Met downstream signalling in the colorectal cancer cell line HCT116.
Analysis of the TCGA colorectal adenocarcinoma data set showed that the expression of Itch was altered in 36% of the patient samples, and Met in 7% of the samples. The prognosis for patients carrying changes in both was worse than for either one alone (p=0.066, N=193). Overexpression of Itch in HCT116 cells did not affect ERK1/2 signalling downstream of the EGFR or Met receptor. However, expression of a dominant negative ligase mutant reduced ERK1/2 signalling from the Met receptor but not the EGF receptor. Thus, suggesting that Itch is important for Met signalling. Further supporting this notion, our knockdown experiments showed a significant effect on HGF-induced but not EGF-induced ERK signalling.
Knockdown of Itch in cultured cells showed aberrant endocytic protein levels, as assessed by Western blot. These proteins are involved in clathrin-mediated endocytosis and their expression levels regulate the speed and efficiency of receptor internalisation, thus implicating a novel Itch-mediated regulatory mechanism for membrane trafficking. Our study highlights the importance of assessing the regulation of receptor signalling and membrane trafficking in colorectal cancer.
Original languageEnglish
Title of host publication2017 Irish Association for Cancer Research: Proceedings
Publication statusPublished - Feb 2017
EventThe Irish Association for Cancer Research - Kilkenny, Ireland
Duration: 22 Feb 201723 Feb 2017

Conference

ConferenceThe Irish Association for Cancer Research
CountryIreland
CityKilkenny
Period22/02/201723/02/2017

Fingerprint

Ubiquitin-Protein Ligases
Receptor Protein-Tyrosine Kinases
Colorectal Neoplasms
Neoplasm Metastasis
Endocytosis
Cell Line
Membranes
HCT116 Cells
Neoplasms
Clathrin
Ubiquitination
Ligases
Epidermal Growth Factor Receptor
Epidermal Growth Factor
Cell Movement
Cultured Cells
Cell Survival
Carcinogenesis
Proteins
Adenocarcinoma

Cite this

@inproceedings{ab5f25aabb6b475cb20ccc1a3ddf71bd,
title = "Itch, an E3 ubiquitin ligase, influences downstream receptor tyrosine kinase signalling in the colorectal cancer cell line, HCT116",
abstract = "Receptor tyrosine kinase signalling is an important driver of cancer cell development. Deregulation of the receptor tyrosine kinases EGFR and Met is associated with cancer formation and progression to metastasis. Recently, mutations in Met that affect endocytic trafficking have been shown to promote cell migration, metastasis and tumorigenesis due to increased endosomal signalling. Thus, Met endocytic trafficking regulates signalling pathways; such as ERK1/2, STAT3, Gab1 and Rac1; that are implicated in cell survival, invasion and metastasis. Advancements in our understanding of regulation of EGFR and Met signalling is essential in order to understand their role in cancer progression. Ubiquitination regulates both receptor degradation and membrane trafficking. Here we investigated the role of the ubiquitin E3 ligase Itch in endocytosis and EGFR and Met downstream signalling in the colorectal cancer cell line HCT116. Analysis of the TCGA colorectal adenocarcinoma data set showed that the expression of Itch was altered in 36{\%} of the patient samples, and Met in 7{\%} of the samples. The prognosis for patients carrying changes in both was worse than for either one alone (p=0.066, N=193). Overexpression of Itch in HCT116 cells did not affect ERK1/2 signalling downstream of the EGFR or Met receptor. However, expression of a dominant negative ligase mutant reduced ERK1/2 signalling from the Met receptor but not the EGF receptor. Thus, suggesting that Itch is important for Met signalling. Further supporting this notion, our knockdown experiments showed a significant effect on HGF-induced but not EGF-induced ERK signalling.Knockdown of Itch in cultured cells showed aberrant endocytic protein levels, as assessed by Western blot. These proteins are involved in clathrin-mediated endocytosis and their expression levels regulate the speed and efficiency of receptor internalisation, thus implicating a novel Itch-mediated regulatory mechanism for membrane trafficking. Our study highlights the importance of assessing the regulation of receptor signalling and membrane trafficking in colorectal cancer.",
author = "Aidan Seeley and Daniel Longley and Emma Evergren",
year = "2017",
month = "2",
language = "English",
booktitle = "2017 Irish Association for Cancer Research: Proceedings",

}

Seeley, A, Longley, D & Evergren, E 2017, Itch, an E3 ubiquitin ligase, influences downstream receptor tyrosine kinase signalling in the colorectal cancer cell line, HCT116. in 2017 Irish Association for Cancer Research: Proceedings. The Irish Association for Cancer Research, Kilkenny, Ireland, 22/02/2017.

Itch, an E3 ubiquitin ligase, influences downstream receptor tyrosine kinase signalling in the colorectal cancer cell line, HCT116. / Seeley, Aidan; Longley, Daniel; Evergren, Emma.

2017 Irish Association for Cancer Research: Proceedings. 2017.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

TY - GEN

T1 - Itch, an E3 ubiquitin ligase, influences downstream receptor tyrosine kinase signalling in the colorectal cancer cell line, HCT116

AU - Seeley, Aidan

AU - Longley, Daniel

AU - Evergren, Emma

PY - 2017/2

Y1 - 2017/2

N2 - Receptor tyrosine kinase signalling is an important driver of cancer cell development. Deregulation of the receptor tyrosine kinases EGFR and Met is associated with cancer formation and progression to metastasis. Recently, mutations in Met that affect endocytic trafficking have been shown to promote cell migration, metastasis and tumorigenesis due to increased endosomal signalling. Thus, Met endocytic trafficking regulates signalling pathways; such as ERK1/2, STAT3, Gab1 and Rac1; that are implicated in cell survival, invasion and metastasis. Advancements in our understanding of regulation of EGFR and Met signalling is essential in order to understand their role in cancer progression. Ubiquitination regulates both receptor degradation and membrane trafficking. Here we investigated the role of the ubiquitin E3 ligase Itch in endocytosis and EGFR and Met downstream signalling in the colorectal cancer cell line HCT116. Analysis of the TCGA colorectal adenocarcinoma data set showed that the expression of Itch was altered in 36% of the patient samples, and Met in 7% of the samples. The prognosis for patients carrying changes in both was worse than for either one alone (p=0.066, N=193). Overexpression of Itch in HCT116 cells did not affect ERK1/2 signalling downstream of the EGFR or Met receptor. However, expression of a dominant negative ligase mutant reduced ERK1/2 signalling from the Met receptor but not the EGF receptor. Thus, suggesting that Itch is important for Met signalling. Further supporting this notion, our knockdown experiments showed a significant effect on HGF-induced but not EGF-induced ERK signalling.Knockdown of Itch in cultured cells showed aberrant endocytic protein levels, as assessed by Western blot. These proteins are involved in clathrin-mediated endocytosis and their expression levels regulate the speed and efficiency of receptor internalisation, thus implicating a novel Itch-mediated regulatory mechanism for membrane trafficking. Our study highlights the importance of assessing the regulation of receptor signalling and membrane trafficking in colorectal cancer.

AB - Receptor tyrosine kinase signalling is an important driver of cancer cell development. Deregulation of the receptor tyrosine kinases EGFR and Met is associated with cancer formation and progression to metastasis. Recently, mutations in Met that affect endocytic trafficking have been shown to promote cell migration, metastasis and tumorigenesis due to increased endosomal signalling. Thus, Met endocytic trafficking regulates signalling pathways; such as ERK1/2, STAT3, Gab1 and Rac1; that are implicated in cell survival, invasion and metastasis. Advancements in our understanding of regulation of EGFR and Met signalling is essential in order to understand their role in cancer progression. Ubiquitination regulates both receptor degradation and membrane trafficking. Here we investigated the role of the ubiquitin E3 ligase Itch in endocytosis and EGFR and Met downstream signalling in the colorectal cancer cell line HCT116. Analysis of the TCGA colorectal adenocarcinoma data set showed that the expression of Itch was altered in 36% of the patient samples, and Met in 7% of the samples. The prognosis for patients carrying changes in both was worse than for either one alone (p=0.066, N=193). Overexpression of Itch in HCT116 cells did not affect ERK1/2 signalling downstream of the EGFR or Met receptor. However, expression of a dominant negative ligase mutant reduced ERK1/2 signalling from the Met receptor but not the EGF receptor. Thus, suggesting that Itch is important for Met signalling. Further supporting this notion, our knockdown experiments showed a significant effect on HGF-induced but not EGF-induced ERK signalling.Knockdown of Itch in cultured cells showed aberrant endocytic protein levels, as assessed by Western blot. These proteins are involved in clathrin-mediated endocytosis and their expression levels regulate the speed and efficiency of receptor internalisation, thus implicating a novel Itch-mediated regulatory mechanism for membrane trafficking. Our study highlights the importance of assessing the regulation of receptor signalling and membrane trafficking in colorectal cancer.

M3 - Conference contribution

BT - 2017 Irish Association for Cancer Research: Proceedings

ER -