Hepatitis B virus X protein (HBx) enhances centrosomal P4.1-associated protein (CPAP) expression to promote hepatocarcinogenesis

Chia Jui Yen, Shu Ting Yang, Ruo Yu Chen, Wenya Huang, Kazuaki Chayama, Ming Hao Lee, Shiang Jie Yang, Hong Sheng Lai, Hsin Yi Yen, Yu Wei Hsiao, Ju Ming Wang, Yih Jyh Lin, Liang Yi Hung

研究成果: Article

摘要

Background: Our previous report suggested that centrosomal P4.1-associated protein (CPAP) is required for Hepatitis B virus (HBV) encoded non-structure protein X (HBx)-mediated nuclear factor kappa light chain enhancer of activated B cells (NF-andkappa;B) activation. CPAP is overexpressed in HBV-associated hepatocellular carcinoma (HCC); however, the interaction between CPAP and HBx in HBV-HCC remains unclear. Methods: The mRNA expression of CPAP and HBx was analyzed by quantitative-PCR (Q-PCR). NF-andkappa;B transcriptional activity and CPAP promoter activity were determined using a reporter assay in Huh7 and Hep3B cells. Immunoprecipitation (IP) and in situ proximal ligation assay (PLA) were performed to detect the interaction between CPAP and HBx. Chromatin-IP was used to detect the association of cAMP response element binding protein (CREB) and HBx with the CPAP promoter. Cell proliferation was measured using cell counting kit CCK-8, Bromodeoxyuridine (5-bromo-2andprime;-deoxyuridine, BrdU) incorporation, and clonogenic assays. The tumorigenic effects of CPAP were determined using xenograft animal models. Results: HBx can transcriptionally up-regulate CPAP via interacting with CREB. Overexpressed CPAP directly interacted with HBx to promote HBx-mediated cell proliferation and migration; SUMO modification of CPAP was involved in interacting with HBx. Knocked-down expression of CPAP decreased the HBx-mediated tumorigenic effects, including cytokines secretion. Interestingly, overexpressed CPAP maintained the HBx protein stability in an NF-andkappa;B-dependent manner; and the expression levels of CPAP and HBx were positively correlated with the activation status of NF-andkappa;B in HCC. Increased expression of CPAP and CREB mRNAs existed in the high-risk group with a lower survival rate in HBV-HCC. Conclusion: The interaction between CPAP and HBx can provide a microenvironment to facilitate HCC development via enhancing NF-andkappa;B activation, inflammatory cytokine production, and cancer malignancies. This study not only sheds light on the role of CPAP in HBV-associated HCC, but also provides CPAP as a potential target for blocking the hyper-activated NF-andkappa;B in HCC. andcopy; 2019 The Author(s).

原文English
文章編號44
期刊Journal of biomedical science
26
發行號1
DOIs
出版狀態Published - 2019 六月 6

指紋

Proteins
Hepatocellular Carcinoma
Viruses
Hepatitis B virus
Cyclic AMP Response Element-Binding Protein
hepatitis B virus X protein
Assays
Chemical activation
Bromodeoxyuridine
Cell proliferation
Cell Proliferation
Cytokines
Sincalide
Deoxyuridine
Messenger RNA
Protein Stability
Chromatin Immunoprecipitation
Immunoprecipitation
Heterografts
Cell Movement

All Science Journal Classification (ASJC) codes

  • Endocrinology, Diabetes and Metabolism
  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology
  • Biochemistry, medical
  • Pharmacology (medical)

引用此文

@article{b2fab636627b4fe795fb255bdd4b25eb,
title = "Hepatitis B virus X protein (HBx) enhances centrosomal P4.1-associated protein (CPAP) expression to promote hepatocarcinogenesis",
abstract = "Background: Our previous report suggested that centrosomal P4.1-associated protein (CPAP) is required for Hepatitis B virus (HBV) encoded non-structure protein X (HBx)-mediated nuclear factor kappa light chain enhancer of activated B cells (NF-andkappa;B) activation. CPAP is overexpressed in HBV-associated hepatocellular carcinoma (HCC); however, the interaction between CPAP and HBx in HBV-HCC remains unclear. Methods: The mRNA expression of CPAP and HBx was analyzed by quantitative-PCR (Q-PCR). NF-andkappa;B transcriptional activity and CPAP promoter activity were determined using a reporter assay in Huh7 and Hep3B cells. Immunoprecipitation (IP) and in situ proximal ligation assay (PLA) were performed to detect the interaction between CPAP and HBx. Chromatin-IP was used to detect the association of cAMP response element binding protein (CREB) and HBx with the CPAP promoter. Cell proliferation was measured using cell counting kit CCK-8, Bromodeoxyuridine (5-bromo-2andprime;-deoxyuridine, BrdU) incorporation, and clonogenic assays. The tumorigenic effects of CPAP were determined using xenograft animal models. Results: HBx can transcriptionally up-regulate CPAP via interacting with CREB. Overexpressed CPAP directly interacted with HBx to promote HBx-mediated cell proliferation and migration; SUMO modification of CPAP was involved in interacting with HBx. Knocked-down expression of CPAP decreased the HBx-mediated tumorigenic effects, including cytokines secretion. Interestingly, overexpressed CPAP maintained the HBx protein stability in an NF-andkappa;B-dependent manner; and the expression levels of CPAP and HBx were positively correlated with the activation status of NF-andkappa;B in HCC. Increased expression of CPAP and CREB mRNAs existed in the high-risk group with a lower survival rate in HBV-HCC. Conclusion: The interaction between CPAP and HBx can provide a microenvironment to facilitate HCC development via enhancing NF-andkappa;B activation, inflammatory cytokine production, and cancer malignancies. This study not only sheds light on the role of CPAP in HBV-associated HCC, but also provides CPAP as a potential target for blocking the hyper-activated NF-andkappa;B in HCC. andcopy; 2019 The Author(s).",
author = "Yen, {Chia Jui} and Yang, {Shu Ting} and Chen, {Ruo Yu} and Wenya Huang and Kazuaki Chayama and Lee, {Ming Hao} and Yang, {Shiang Jie} and Lai, {Hong Sheng} and Yen, {Hsin Yi} and Hsiao, {Yu Wei} and Wang, {Ju Ming} and Lin, {Yih Jyh} and Hung, {Liang Yi}",
year = "2019",
month = "6",
day = "6",
doi = "10.1186/s12929-019-0534-9",
language = "English",
volume = "26",
journal = "Journal of Biomedical Science",
issn = "1021-7770",
publisher = "BioMed Central",
number = "1",

}

Hepatitis B virus X protein (HBx) enhances centrosomal P4.1-associated protein (CPAP) expression to promote hepatocarcinogenesis. / Yen, Chia Jui; Yang, Shu Ting; Chen, Ruo Yu; Huang, Wenya; Chayama, Kazuaki; Lee, Ming Hao; Yang, Shiang Jie; Lai, Hong Sheng; Yen, Hsin Yi; Hsiao, Yu Wei; Wang, Ju Ming; Lin, Yih Jyh; Hung, Liang Yi.

於: Journal of biomedical science, 卷 26, 編號 1, 44, 06.06.2019.

研究成果: Article

TY - JOUR

T1 - Hepatitis B virus X protein (HBx) enhances centrosomal P4.1-associated protein (CPAP) expression to promote hepatocarcinogenesis

AU - Yen, Chia Jui

AU - Yang, Shu Ting

AU - Chen, Ruo Yu

AU - Huang, Wenya

AU - Chayama, Kazuaki

AU - Lee, Ming Hao

AU - Yang, Shiang Jie

AU - Lai, Hong Sheng

AU - Yen, Hsin Yi

AU - Hsiao, Yu Wei

AU - Wang, Ju Ming

AU - Lin, Yih Jyh

AU - Hung, Liang Yi

PY - 2019/6/6

Y1 - 2019/6/6

N2 - Background: Our previous report suggested that centrosomal P4.1-associated protein (CPAP) is required for Hepatitis B virus (HBV) encoded non-structure protein X (HBx)-mediated nuclear factor kappa light chain enhancer of activated B cells (NF-andkappa;B) activation. CPAP is overexpressed in HBV-associated hepatocellular carcinoma (HCC); however, the interaction between CPAP and HBx in HBV-HCC remains unclear. Methods: The mRNA expression of CPAP and HBx was analyzed by quantitative-PCR (Q-PCR). NF-andkappa;B transcriptional activity and CPAP promoter activity were determined using a reporter assay in Huh7 and Hep3B cells. Immunoprecipitation (IP) and in situ proximal ligation assay (PLA) were performed to detect the interaction between CPAP and HBx. Chromatin-IP was used to detect the association of cAMP response element binding protein (CREB) and HBx with the CPAP promoter. Cell proliferation was measured using cell counting kit CCK-8, Bromodeoxyuridine (5-bromo-2andprime;-deoxyuridine, BrdU) incorporation, and clonogenic assays. The tumorigenic effects of CPAP were determined using xenograft animal models. Results: HBx can transcriptionally up-regulate CPAP via interacting with CREB. Overexpressed CPAP directly interacted with HBx to promote HBx-mediated cell proliferation and migration; SUMO modification of CPAP was involved in interacting with HBx. Knocked-down expression of CPAP decreased the HBx-mediated tumorigenic effects, including cytokines secretion. Interestingly, overexpressed CPAP maintained the HBx protein stability in an NF-andkappa;B-dependent manner; and the expression levels of CPAP and HBx were positively correlated with the activation status of NF-andkappa;B in HCC. Increased expression of CPAP and CREB mRNAs existed in the high-risk group with a lower survival rate in HBV-HCC. Conclusion: The interaction between CPAP and HBx can provide a microenvironment to facilitate HCC development via enhancing NF-andkappa;B activation, inflammatory cytokine production, and cancer malignancies. This study not only sheds light on the role of CPAP in HBV-associated HCC, but also provides CPAP as a potential target for blocking the hyper-activated NF-andkappa;B in HCC. andcopy; 2019 The Author(s).

AB - Background: Our previous report suggested that centrosomal P4.1-associated protein (CPAP) is required for Hepatitis B virus (HBV) encoded non-structure protein X (HBx)-mediated nuclear factor kappa light chain enhancer of activated B cells (NF-andkappa;B) activation. CPAP is overexpressed in HBV-associated hepatocellular carcinoma (HCC); however, the interaction between CPAP and HBx in HBV-HCC remains unclear. Methods: The mRNA expression of CPAP and HBx was analyzed by quantitative-PCR (Q-PCR). NF-andkappa;B transcriptional activity and CPAP promoter activity were determined using a reporter assay in Huh7 and Hep3B cells. Immunoprecipitation (IP) and in situ proximal ligation assay (PLA) were performed to detect the interaction between CPAP and HBx. Chromatin-IP was used to detect the association of cAMP response element binding protein (CREB) and HBx with the CPAP promoter. Cell proliferation was measured using cell counting kit CCK-8, Bromodeoxyuridine (5-bromo-2andprime;-deoxyuridine, BrdU) incorporation, and clonogenic assays. The tumorigenic effects of CPAP were determined using xenograft animal models. Results: HBx can transcriptionally up-regulate CPAP via interacting with CREB. Overexpressed CPAP directly interacted with HBx to promote HBx-mediated cell proliferation and migration; SUMO modification of CPAP was involved in interacting with HBx. Knocked-down expression of CPAP decreased the HBx-mediated tumorigenic effects, including cytokines secretion. Interestingly, overexpressed CPAP maintained the HBx protein stability in an NF-andkappa;B-dependent manner; and the expression levels of CPAP and HBx were positively correlated with the activation status of NF-andkappa;B in HCC. Increased expression of CPAP and CREB mRNAs existed in the high-risk group with a lower survival rate in HBV-HCC. Conclusion: The interaction between CPAP and HBx can provide a microenvironment to facilitate HCC development via enhancing NF-andkappa;B activation, inflammatory cytokine production, and cancer malignancies. This study not only sheds light on the role of CPAP in HBV-associated HCC, but also provides CPAP as a potential target for blocking the hyper-activated NF-andkappa;B in HCC. andcopy; 2019 The Author(s).

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U2 - 10.1186/s12929-019-0534-9

DO - 10.1186/s12929-019-0534-9

M3 - Article

C2 - 31170980

AN - SCOPUS:85066961895

VL - 26

JO - Journal of Biomedical Science

JF - Journal of Biomedical Science

SN - 1021-7770

IS - 1

M1 - 44

ER -