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Hepatitis B Virus Core Promoter A1762T/G1764A (TA)/T1753A/T1768A Mutations Contribute to Hepatocarcinogenesis by Deregulating Skp2 and P53

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Abstract

Background and Aim

Hepatitis B virus core promoter (CP) mutations can increase risk of hepatocellular carcinoma. The CP region overlaps with the HBV X (HBx) gene, which has been associated with hepatocarcinogenesis. The cyclin kinase inhibitor P53 is an important regulator of cell cycle progression. We determined whether HBx mutants that result from mutations in the CP deregulate P53.

Methods

A HBx combination (combo) mutant with changes in the CP region that corresponded to A1762T/G1764A (TA), T1753A, and T1768A was constructed and expressed in L-02 and Hep3B cells. The effects of CP mutations on expression and degradation of P53, and the effects on cell cycle progression and proliferation were analyzed.

Results

The combo mutant decreased levels of P53 and increased cyclin D1 expression, accelerated P53 degradation in L-02 cells, accelerated cell cycle progression, and increased expression of S-phase kinase-associated protein 2 (Skp2) in L-02 and Hep3B cells. Silencing of Skp2 abrogated the effects of CP mutations on P53 expression. The kinetics of P53 expression correlated with changes in cell cycle distribution.

Conclusions

The HBx mutant with a combination of CP mutations can up-regulate Skp2, which then down-regulates P53 via ubiquitin-mediated proteasomal degradation, increasing the risk of hepatocellular carcinoma.

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Acknowledgments

This work was supported by National Natural Science Foundation of Guangdong Province (S2013010016015, Meihai Deng), The National Natural Science Fund for Young Scholars of China (81000177, Yuesi Zhong) and The Projects on Science and Technology plan of Guangzhou City (2013J410061-1955, Meihai Deng).

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Authors and Affiliations

  1. Hepatobiliary Surgery Department, The Third Affiliated Hospital of Sun Yat-Sen University, No. 600, TianHe Road, TianHe District, Guangzhou City, 510630, China

    Jian Yan, Yuesi Zhong, Zhiyong Xiong & Meihai Deng

  2. Department of General Surgery, Lingnan Hospital of Sun Yat-Sen University, No. 600, TianHe Road, TianHe District, Guangzhou City, 510630, China

    Zhicheng Yao & Kunpeng Hu

  3. Department of Intensive Care Unit, Lingnan Hospital of Sun Yat-Sen University, No. 600, TianHe Road, TianHe District, Guangzhou City, 510630, China

    Mingliang Li

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  1. Jian Yan
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  2. Zhicheng Yao
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  3. Kunpeng Hu
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  4. Yuesi Zhong
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Correspondence to Meihai Deng.

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Zhicheng Yao and Kunpeng Hu have contributed equally to this work.

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Yan, J., Yao, Z., Hu, K. et al. Hepatitis B Virus Core Promoter A1762T/G1764A (TA)/T1753A/T1768A Mutations Contribute to Hepatocarcinogenesis by Deregulating Skp2 and P53. Dig Dis Sci 60, 1315–1324 (2015). https://doi.org/10.1007/s10620-014-3492-9

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