SUMOylated CPAP is required for IKK-mediated NF-κB activation and enhances HBx-induced NF-κB signaling in HCC

Shu Ting Yang, Chia Jui Yen, Chien Hsien Lai, Yih Jyh Lin, Kung Chao Chang, Jenq Chang Lee, Yao Wen Liu, Pey Yi Chang-Liao, Lu Shin Hsu, Wen Chang Chang, Wen Chun Hung, Tang K. Tang, Yi Wen Liu, Liang Yi Hung

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Abstract

Background & Aims: Constitutive activation of NF-κB is an important event involved in chronic inflammation in hepatocellular carcinoma (HCC). CPAP, which plays important roles in centrosomal functions, was previously identified as the transcriptional co-activator of NF-κB. However, the molecular mechanism is unclear. The goal of this study was to investigate the role of CPAP in activating the NF-κB pathway in HCC. Methods: SK-Hep1, HuH7, HepG2, HepG2X, Hep3B, and Hep3BX cells with CPAP overexpression or CPAP siRNA were used to evaluate activation of NF-κB under TNF-α stimulation by reporter assay, RT-PCR, Q-PCR, and Western blot analysis. In vivo SUMO modification of CPAP was demonstrated by an in situ PLA assay. Human HCC tissues were used to perform Q-PCR, Western blot, and IHC. Results: CPAP siRNA abolished the interaction between IKKβ and NF-κB, whereas overexpression of CPAP enhanced this interaction and finally led to augmented NF-κB activation by increasing the phosphorylation of NF-κB. CPAP could enter nuclei by associating with NF-κB. Furthermore, CPAP was SUMO-1 modified upon TNF-α stimulus, and this is essential for its NF-κB co-activator activity. SUMO-1-deficient CPAP mutant lost its NF-κB co-activator activity and failed to enter nuclei. Importantly, SUMOylated CPAP could synergistically increase the HBx-induced NF-κB activity. Conclusions: CPAP is essential for the recruitment of the IKK complex to inactivated NF-κB upon TNF-α treatment. Expression of CPAP was positively correlated with a poor prognosis in HBV-HCC. CPAP has the potential to serve as a therapeutic target for inflammation and inflammation-related diseases.

Original languageEnglish
Pages (from-to)1157-1164
Number of pages8
JournalJournal of Hepatology
Volume58
Issue number6
DOIs
Publication statusPublished - 2013 Jun 1

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Hepatocellular Carcinoma
Inflammation
Polymerase Chain Reaction
Small Interfering RNA
Western Blotting
Phosphorylation
Therapeutics

All Science Journal Classification (ASJC) codes

  • Hepatology

Cite this

Yang, Shu Ting ; Yen, Chia Jui ; Lai, Chien Hsien ; Lin, Yih Jyh ; Chang, Kung Chao ; Lee, Jenq Chang ; Liu, Yao Wen ; Chang-Liao, Pey Yi ; Hsu, Lu Shin ; Chang, Wen Chang ; Hung, Wen Chun ; Tang, Tang K. ; Liu, Yi Wen ; Hung, Liang Yi. / SUMOylated CPAP is required for IKK-mediated NF-κB activation and enhances HBx-induced NF-κB signaling in HCC. In: Journal of Hepatology. 2013 ; Vol. 58, No. 6. pp. 1157-1164.
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title = "SUMOylated CPAP is required for IKK-mediated NF-κB activation and enhances HBx-induced NF-κB signaling in HCC",
abstract = "Background & Aims: Constitutive activation of NF-κB is an important event involved in chronic inflammation in hepatocellular carcinoma (HCC). CPAP, which plays important roles in centrosomal functions, was previously identified as the transcriptional co-activator of NF-κB. However, the molecular mechanism is unclear. The goal of this study was to investigate the role of CPAP in activating the NF-κB pathway in HCC. Methods: SK-Hep1, HuH7, HepG2, HepG2X, Hep3B, and Hep3BX cells with CPAP overexpression or CPAP siRNA were used to evaluate activation of NF-κB under TNF-α stimulation by reporter assay, RT-PCR, Q-PCR, and Western blot analysis. In vivo SUMO modification of CPAP was demonstrated by an in situ PLA assay. Human HCC tissues were used to perform Q-PCR, Western blot, and IHC. Results: CPAP siRNA abolished the interaction between IKKβ and NF-κB, whereas overexpression of CPAP enhanced this interaction and finally led to augmented NF-κB activation by increasing the phosphorylation of NF-κB. CPAP could enter nuclei by associating with NF-κB. Furthermore, CPAP was SUMO-1 modified upon TNF-α stimulus, and this is essential for its NF-κB co-activator activity. SUMO-1-deficient CPAP mutant lost its NF-κB co-activator activity and failed to enter nuclei. Importantly, SUMOylated CPAP could synergistically increase the HBx-induced NF-κB activity. Conclusions: CPAP is essential for the recruitment of the IKK complex to inactivated NF-κB upon TNF-α treatment. Expression of CPAP was positively correlated with a poor prognosis in HBV-HCC. CPAP has the potential to serve as a therapeutic target for inflammation and inflammation-related diseases.",
author = "Yang, {Shu Ting} and Yen, {Chia Jui} and Lai, {Chien Hsien} and Lin, {Yih Jyh} and Chang, {Kung Chao} and Lee, {Jenq Chang} and Liu, {Yao Wen} and Chang-Liao, {Pey Yi} and Hsu, {Lu Shin} and Chang, {Wen Chang} and Hung, {Wen Chun} and Tang, {Tang K.} and Liu, {Yi Wen} and Hung, {Liang Yi}",
year = "2013",
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SUMOylated CPAP is required for IKK-mediated NF-κB activation and enhances HBx-induced NF-κB signaling in HCC. / Yang, Shu Ting; Yen, Chia Jui; Lai, Chien Hsien; Lin, Yih Jyh; Chang, Kung Chao; Lee, Jenq Chang; Liu, Yao Wen; Chang-Liao, Pey Yi; Hsu, Lu Shin; Chang, Wen Chang; Hung, Wen Chun; Tang, Tang K.; Liu, Yi Wen; Hung, Liang Yi.

In: Journal of Hepatology, Vol. 58, No. 6, 01.06.2013, p. 1157-1164.

Research output: Contribution to journalArticle

TY - JOUR

T1 - SUMOylated CPAP is required for IKK-mediated NF-κB activation and enhances HBx-induced NF-κB signaling in HCC

AU - Yang, Shu Ting

AU - Yen, Chia Jui

AU - Lai, Chien Hsien

AU - Lin, Yih Jyh

AU - Chang, Kung Chao

AU - Lee, Jenq Chang

AU - Liu, Yao Wen

AU - Chang-Liao, Pey Yi

AU - Hsu, Lu Shin

AU - Chang, Wen Chang

AU - Hung, Wen Chun

AU - Tang, Tang K.

AU - Liu, Yi Wen

AU - Hung, Liang Yi

PY - 2013/6/1

Y1 - 2013/6/1

N2 - Background & Aims: Constitutive activation of NF-κB is an important event involved in chronic inflammation in hepatocellular carcinoma (HCC). CPAP, which plays important roles in centrosomal functions, was previously identified as the transcriptional co-activator of NF-κB. However, the molecular mechanism is unclear. The goal of this study was to investigate the role of CPAP in activating the NF-κB pathway in HCC. Methods: SK-Hep1, HuH7, HepG2, HepG2X, Hep3B, and Hep3BX cells with CPAP overexpression or CPAP siRNA were used to evaluate activation of NF-κB under TNF-α stimulation by reporter assay, RT-PCR, Q-PCR, and Western blot analysis. In vivo SUMO modification of CPAP was demonstrated by an in situ PLA assay. Human HCC tissues were used to perform Q-PCR, Western blot, and IHC. Results: CPAP siRNA abolished the interaction between IKKβ and NF-κB, whereas overexpression of CPAP enhanced this interaction and finally led to augmented NF-κB activation by increasing the phosphorylation of NF-κB. CPAP could enter nuclei by associating with NF-κB. Furthermore, CPAP was SUMO-1 modified upon TNF-α stimulus, and this is essential for its NF-κB co-activator activity. SUMO-1-deficient CPAP mutant lost its NF-κB co-activator activity and failed to enter nuclei. Importantly, SUMOylated CPAP could synergistically increase the HBx-induced NF-κB activity. Conclusions: CPAP is essential for the recruitment of the IKK complex to inactivated NF-κB upon TNF-α treatment. Expression of CPAP was positively correlated with a poor prognosis in HBV-HCC. CPAP has the potential to serve as a therapeutic target for inflammation and inflammation-related diseases.

AB - Background & Aims: Constitutive activation of NF-κB is an important event involved in chronic inflammation in hepatocellular carcinoma (HCC). CPAP, which plays important roles in centrosomal functions, was previously identified as the transcriptional co-activator of NF-κB. However, the molecular mechanism is unclear. The goal of this study was to investigate the role of CPAP in activating the NF-κB pathway in HCC. Methods: SK-Hep1, HuH7, HepG2, HepG2X, Hep3B, and Hep3BX cells with CPAP overexpression or CPAP siRNA were used to evaluate activation of NF-κB under TNF-α stimulation by reporter assay, RT-PCR, Q-PCR, and Western blot analysis. In vivo SUMO modification of CPAP was demonstrated by an in situ PLA assay. Human HCC tissues were used to perform Q-PCR, Western blot, and IHC. Results: CPAP siRNA abolished the interaction between IKKβ and NF-κB, whereas overexpression of CPAP enhanced this interaction and finally led to augmented NF-κB activation by increasing the phosphorylation of NF-κB. CPAP could enter nuclei by associating with NF-κB. Furthermore, CPAP was SUMO-1 modified upon TNF-α stimulus, and this is essential for its NF-κB co-activator activity. SUMO-1-deficient CPAP mutant lost its NF-κB co-activator activity and failed to enter nuclei. Importantly, SUMOylated CPAP could synergistically increase the HBx-induced NF-κB activity. Conclusions: CPAP is essential for the recruitment of the IKK complex to inactivated NF-κB upon TNF-α treatment. Expression of CPAP was positively correlated with a poor prognosis in HBV-HCC. CPAP has the potential to serve as a therapeutic target for inflammation and inflammation-related diseases.

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U2 - 10.1016/j.jhep.2013.01.025

DO - 10.1016/j.jhep.2013.01.025

M3 - Article

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JO - Journal of Hepatology

JF - Journal of Hepatology

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