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HBx interacted with Smad4 to deprive activin a growth inhibition function in hepatocyte HL7702 on CRM1 manner

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Tumor Biology

Abstract

Hepatitis B virus (HBV) is implicated in the pathogenesis of hepatocellular carcinoma, which has been found to be associated with TGF-beta signaling. Activin A is a TGF-β family cytokine that exhibits cell proliferation inhibition on normal hepatocyte. How HBV-encoded X oncoprotein play in activin’s activity on hepatocyte has not been developed. In this study, a nontumor hepatic cell line HL7702 with HBX ectogenic expression has been established. MTT and BrdU assays showed that HBx promoted growth of HL7702 cells in vitro and downregulated activin signaling. Deregulated activin signaling pathway by HBX failed to activate target gene p21/waf1 and p15 transcription. In addition, mammalian two-hybrid and coimmunoprecipitation assays revealed that HBX could directly interact with activin signaling transduction protein Smad4, making activated Smad2/3/4 nucleus translocation suppressed. Furthermore, we detected that leptomycin B, the inhibitor of CRM1 protein, could recover nuclear translocation of endogenous Smads complex in HL7702 with HBX expression, indicating that HBX antagonized Smads nucleus translocation, at least partially, on CRM1-dependent manner. Leptomycin B was found to have antigrowth activity on HBX-expressed HL7702, according to its antitumor function in previous study. Above all, HBX antagonized activin signaling in normal human liver cells by interacting with Smad4 might one of the considerable causes of HBX-induced hepatocyte transformation, which deprived activin’s cell growth inhibition function at an early stage of tumorigenesis.

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Acknowledgements

This study was supported by grants from the National Natural Science Foundation of China (81301414) and the China Postdoctoral Science Foundation (2013M540245, 2014T70283) to Mingyu Lv, and The Fourth Youth Science Foundation of First Hospital of Jilin University (20121224) to Ying Shi.

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Authors and Affiliations

  1. Institute of Translational Medicine, First Hospital of Jilin University, Changchun, China

    Ying Shi, Zhu Han, Wenyan Zhang & Mingyu Lv

  2. The First Clinical Medical College of Jilin University, Changchun, China

    Haipeng Zhang

  3. Tumor Biological Treatment Center of Jilin Province People’s Hospital, Changchun, China

    Xuguang Mi

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  1. Ying Shi
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  2. Haipeng Zhang
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  3. Zhu Han
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  4. Xuguang Mi
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  5. Wenyan Zhang
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Correspondence to Wenyan Zhang or Mingyu Lv.

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Shi, Y., Zhang, H., Han, Z. et al. HBx interacted with Smad4 to deprive activin a growth inhibition function in hepatocyte HL7702 on CRM1 manner. Tumor Biol. 37, 3405–3415 (2016). https://doi.org/10.1007/s13277-015-4076-9

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  • DOI: https://doi.org/10.1007/s13277-015-4076-9

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