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, Wen-Ya 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

Research output: Contribution to journalArticle

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).

Original languageEnglish
Article number44
JournalJournal of biomedical science
Volume26
Issue number1
DOIs
Publication statusPublished - 2019 Jun 6

Fingerprint

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)

Cite this

Yen, Chia-Jui ; Yang, Shu Ting ; Chen, Ruo Yu ; Huang, Wen-Ya ; 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. / Hepatitis B virus X protein (HBx) enhances centrosomal P4.1-associated protein (CPAP) expression to promote hepatocarcinogenesis. In: Journal of biomedical science. 2019 ; Vol. 26, No. 1.
@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 = "Chia-Jui Yen and Yang, {Shu Ting} and Chen, {Ruo Yu} and Wen-Ya 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 Ju-Ming Wang and Yih-Jyh Lin and Liang-Yi Hung",
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, Wen-Ya; 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.

In: Journal of biomedical science, Vol. 26, No. 1, 44, 06.06.2019.

Research output: Contribution to journalArticle

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, Wen-Ya

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).

UR - http://www.scopus.com/inward/record.url?scp=85066961895&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85066961895&partnerID=8YFLogxK

U2 - 10.1186/s12929-019-0534-9

DO - 10.1186/s12929-019-0534-9

M3 - Article

VL - 26

JO - Journal of Biomedical Science

JF - Journal of Biomedical Science

SN - 1021-7770

IS - 1

M1 - 44

ER -