Genetic disruption of KEAP1/CUL3 E3 ubiquitin ligase complex components is a key mechanism of NF-KappaB pathway activation in lung cancer

Kelsie L. Thu, Larissa A. Pikor, Raj Chari, Ian M. Wilson, Calum E. MacAulay, John C. English, Ming Sound Tsao, Adi F. Gazdar, Stephen Lam, Wan L. Lam, William W. Lockwood

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta (IKBKB) (IKK-β/IKK-2), which activates NF-κB, is a substrate of the KEAP1-CUL3-RBX1 E3-ubiquitin ligase complex, implicating this complex in NF-κB pathway regulation. We investigated complex component gene disruption as a novel genetic mechanism of NF-κB activation in non-small cell lung cancer. Methods: A total of 644 tumor- and 90 cell-line genomes were analyzed for gene dosage status of the individual complex components and IKBKB. Gene expression of these genes and NF-κB target genes were analyzed in 48 tumors. IKBKB protein levels were assessed in tumors with and without complex or IKBKB genetic disruption. Complex component knockdown was performed to assess effects of the E3-ligase complex on IKBKB and NF-κB levels, and phenotypic importance of IKBKB expression was measured by pharmacological inhibition. Results: We observed strikingly frequent genetic disruption (42%) and aberrant expression (63%) of the E3-ligase complex and IKBKB in the samples examined. Although both adenocarcinomas and squamous cell carcinomas showed complex disruption, the patterns of gene disruption differed. IKBKB levels were elevated with complex disruption, knockdown of complex components increased activated forms of IKBKB and NF-κB proteins, and IKBKB inhibition detriments cell viability, highlighting the biological significance of complex disruption. NF-κB target genes were overexpressed in samples with complex disruption, further demonstrating the effect of complex disruption on NF-κB activity. Conclusions: Gene dosage alteration is a prominent mechanism that disrupts each component of the KEAP1-CUL3-RBX1 complex and its NF-κB stimulating substrate, IKBKB. Herein, we show that, multiple component disruption of this complex represents a novel mechanism of NF-κB activation in non-small cell lung cancer.

Original languageEnglish (US)
Pages (from-to)1521-1529
Number of pages9
JournalJournal of Thoracic Oncology
Volume6
Issue number9
DOIs
StatePublished - Sep 2011

Fingerprint

I-kappa B Kinase
Ubiquitin-Protein Ligases
NF-kappa B
Lung Neoplasms
Gene Dosage
Genes
Non-Small Cell Lung Carcinoma
Gene Components
Tumor Cell Line

Keywords

  • CUL3
  • Genetic disruption
  • IKBKB
  • KEAP1
  • NF-κB signaling
  • RBX1

ASJC Scopus subject areas

  • Oncology
  • Pulmonary and Respiratory Medicine

Cite this

Genetic disruption of KEAP1/CUL3 E3 ubiquitin ligase complex components is a key mechanism of NF-KappaB pathway activation in lung cancer. / Thu, Kelsie L.; Pikor, Larissa A.; Chari, Raj; Wilson, Ian M.; MacAulay, Calum E.; English, John C.; Tsao, Ming Sound; Gazdar, Adi F.; Lam, Stephen; Lam, Wan L.; Lockwood, William W.

In: Journal of Thoracic Oncology, Vol. 6, No. 9, 09.2011, p. 1521-1529.

Research output: Contribution to journalArticle

Thu, KL, Pikor, LA, Chari, R, Wilson, IM, MacAulay, CE, English, JC, Tsao, MS, Gazdar, AF, Lam, S, Lam, WL & Lockwood, WW 2011, 'Genetic disruption of KEAP1/CUL3 E3 ubiquitin ligase complex components is a key mechanism of NF-KappaB pathway activation in lung cancer', Journal of Thoracic Oncology, vol. 6, no. 9, pp. 1521-1529. https://doi.org/10.1097/JTO.0b013e3182289479
Thu, Kelsie L. ; Pikor, Larissa A. ; Chari, Raj ; Wilson, Ian M. ; MacAulay, Calum E. ; English, John C. ; Tsao, Ming Sound ; Gazdar, Adi F. ; Lam, Stephen ; Lam, Wan L. ; Lockwood, William W. / Genetic disruption of KEAP1/CUL3 E3 ubiquitin ligase complex components is a key mechanism of NF-KappaB pathway activation in lung cancer. In: Journal of Thoracic Oncology. 2011 ; Vol. 6, No. 9. pp. 1521-1529.
@article{8b2fb5bb887346e6b75814a427007317,
title = "Genetic disruption of KEAP1/CUL3 E3 ubiquitin ligase complex components is a key mechanism of NF-KappaB pathway activation in lung cancer",
abstract = "Inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta (IKBKB) (IKK-β/IKK-2), which activates NF-κB, is a substrate of the KEAP1-CUL3-RBX1 E3-ubiquitin ligase complex, implicating this complex in NF-κB pathway regulation. We investigated complex component gene disruption as a novel genetic mechanism of NF-κB activation in non-small cell lung cancer. Methods: A total of 644 tumor- and 90 cell-line genomes were analyzed for gene dosage status of the individual complex components and IKBKB. Gene expression of these genes and NF-κB target genes were analyzed in 48 tumors. IKBKB protein levels were assessed in tumors with and without complex or IKBKB genetic disruption. Complex component knockdown was performed to assess effects of the E3-ligase complex on IKBKB and NF-κB levels, and phenotypic importance of IKBKB expression was measured by pharmacological inhibition. Results: We observed strikingly frequent genetic disruption (42{\%}) and aberrant expression (63{\%}) of the E3-ligase complex and IKBKB in the samples examined. Although both adenocarcinomas and squamous cell carcinomas showed complex disruption, the patterns of gene disruption differed. IKBKB levels were elevated with complex disruption, knockdown of complex components increased activated forms of IKBKB and NF-κB proteins, and IKBKB inhibition detriments cell viability, highlighting the biological significance of complex disruption. NF-κB target genes were overexpressed in samples with complex disruption, further demonstrating the effect of complex disruption on NF-κB activity. Conclusions: Gene dosage alteration is a prominent mechanism that disrupts each component of the KEAP1-CUL3-RBX1 complex and its NF-κB stimulating substrate, IKBKB. Herein, we show that, multiple component disruption of this complex represents a novel mechanism of NF-κB activation in non-small cell lung cancer.",
keywords = "CUL3, Genetic disruption, IKBKB, KEAP1, NF-κB signaling, RBX1",
author = "Thu, {Kelsie L.} and Pikor, {Larissa A.} and Raj Chari and Wilson, {Ian M.} and MacAulay, {Calum E.} and English, {John C.} and Tsao, {Ming Sound} and Gazdar, {Adi F.} and Stephen Lam and Lam, {Wan L.} and Lockwood, {William W.}",
year = "2011",
month = "9",
doi = "10.1097/JTO.0b013e3182289479",
language = "English (US)",
volume = "6",
pages = "1521--1529",
journal = "Journal of Thoracic Oncology",
issn = "1556-0864",
publisher = "International Association for the Study of Lung Cancer",
number = "9",

}

TY - JOUR

T1 - Genetic disruption of KEAP1/CUL3 E3 ubiquitin ligase complex components is a key mechanism of NF-KappaB pathway activation in lung cancer

AU - Thu, Kelsie L.

AU - Pikor, Larissa A.

AU - Chari, Raj

AU - Wilson, Ian M.

AU - MacAulay, Calum E.

AU - English, John C.

AU - Tsao, Ming Sound

AU - Gazdar, Adi F.

AU - Lam, Stephen

AU - Lam, Wan L.

AU - Lockwood, William W.

PY - 2011/9

Y1 - 2011/9

N2 - Inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta (IKBKB) (IKK-β/IKK-2), which activates NF-κB, is a substrate of the KEAP1-CUL3-RBX1 E3-ubiquitin ligase complex, implicating this complex in NF-κB pathway regulation. We investigated complex component gene disruption as a novel genetic mechanism of NF-κB activation in non-small cell lung cancer. Methods: A total of 644 tumor- and 90 cell-line genomes were analyzed for gene dosage status of the individual complex components and IKBKB. Gene expression of these genes and NF-κB target genes were analyzed in 48 tumors. IKBKB protein levels were assessed in tumors with and without complex or IKBKB genetic disruption. Complex component knockdown was performed to assess effects of the E3-ligase complex on IKBKB and NF-κB levels, and phenotypic importance of IKBKB expression was measured by pharmacological inhibition. Results: We observed strikingly frequent genetic disruption (42%) and aberrant expression (63%) of the E3-ligase complex and IKBKB in the samples examined. Although both adenocarcinomas and squamous cell carcinomas showed complex disruption, the patterns of gene disruption differed. IKBKB levels were elevated with complex disruption, knockdown of complex components increased activated forms of IKBKB and NF-κB proteins, and IKBKB inhibition detriments cell viability, highlighting the biological significance of complex disruption. NF-κB target genes were overexpressed in samples with complex disruption, further demonstrating the effect of complex disruption on NF-κB activity. Conclusions: Gene dosage alteration is a prominent mechanism that disrupts each component of the KEAP1-CUL3-RBX1 complex and its NF-κB stimulating substrate, IKBKB. Herein, we show that, multiple component disruption of this complex represents a novel mechanism of NF-κB activation in non-small cell lung cancer.

AB - Inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta (IKBKB) (IKK-β/IKK-2), which activates NF-κB, is a substrate of the KEAP1-CUL3-RBX1 E3-ubiquitin ligase complex, implicating this complex in NF-κB pathway regulation. We investigated complex component gene disruption as a novel genetic mechanism of NF-κB activation in non-small cell lung cancer. Methods: A total of 644 tumor- and 90 cell-line genomes were analyzed for gene dosage status of the individual complex components and IKBKB. Gene expression of these genes and NF-κB target genes were analyzed in 48 tumors. IKBKB protein levels were assessed in tumors with and without complex or IKBKB genetic disruption. Complex component knockdown was performed to assess effects of the E3-ligase complex on IKBKB and NF-κB levels, and phenotypic importance of IKBKB expression was measured by pharmacological inhibition. Results: We observed strikingly frequent genetic disruption (42%) and aberrant expression (63%) of the E3-ligase complex and IKBKB in the samples examined. Although both adenocarcinomas and squamous cell carcinomas showed complex disruption, the patterns of gene disruption differed. IKBKB levels were elevated with complex disruption, knockdown of complex components increased activated forms of IKBKB and NF-κB proteins, and IKBKB inhibition detriments cell viability, highlighting the biological significance of complex disruption. NF-κB target genes were overexpressed in samples with complex disruption, further demonstrating the effect of complex disruption on NF-κB activity. Conclusions: Gene dosage alteration is a prominent mechanism that disrupts each component of the KEAP1-CUL3-RBX1 complex and its NF-κB stimulating substrate, IKBKB. Herein, we show that, multiple component disruption of this complex represents a novel mechanism of NF-κB activation in non-small cell lung cancer.

KW - CUL3

KW - Genetic disruption

KW - IKBKB

KW - KEAP1

KW - NF-κB signaling

KW - RBX1

UR - http://www.scopus.com/inward/record.url?scp=80052268120&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80052268120&partnerID=8YFLogxK

U2 - 10.1097/JTO.0b013e3182289479

DO - 10.1097/JTO.0b013e3182289479

M3 - Article

C2 - 21795997

AN - SCOPUS:80052268120

VL - 6

SP - 1521

EP - 1529

JO - Journal of Thoracic Oncology

JF - Journal of Thoracic Oncology

SN - 1556-0864

IS - 9

ER -