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Reactive oxygen species-mediated PKC and integrin signaling promotes tumor progression of human hepatoma HepG2

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Abstract

The poor prognosis and recurrence of HCC are majorly caused by intrahepatic metastasis. Delineating the molecular pathways mediating these processes may benefit developing effective targeting therapies. Using human hepatoma HepG2 as a model, we have found reactive oxygen species (ROS) may cooperate with protein kinase C (PKC) for sustained ERK phosphorylation and migration of HepG2 induced by 12-O-tetradecanoyl-phorbol-13-acetate (TPA). We further investigated whether integrin signaling is involved. Various antagonists of integrin signaling prevented TPA-induced activation of ERK and PKC, ROS generation and migration of HepG2. On the other hand, TPA-induced phosphorylation of integrin signaling components including focal adhesion kinase (FAK), Src (Tyr416) and paxillin (Tyr31 and Ser178) can be prevented by PKC inhibitor Bisindolylmaleimides (BIS) and antioxidant dithiotheritol (DTT). HepG2 overexpressing PKCα contained elevated phosphorylated paxillin. Also, ROS generator phenazine methosulfate and tert-Butyl hydroperoxide may induce phosphorylation of paxillin and activation of PKC. Taken together, ROS mediated cross talk of PKC and integrin for migration of HepG2 induced by TPA. Furthermore, TPA induced intrahepatic metastasis of HepG2 in SCID mice, which was prevented by BIS or (BIS plus DTT). Elevated phosphorylation of paxillin was observed in tumor of mice treated with TPA as compared with those co-treated with TPA/BIS. In summary, the signal pathways for tumor progression of hepatoma induced by TPA can be established both in vitro and in vivo.

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Abbreviations

TPA:

Tetradecanoyl phorbol acetate

PKC:

Protein kinase C

ROS:

Reactive oxygen species

FAK:

Focal adhesion kinase

ERK:

Extracellular signal-regulated kinases

BIS:

Bisindolylmaleimides

DTT:

Dithiotheritol

TBHP:

Tert-butyl hydroperoxide

PMS:

Phenazine methosulfate

HCC:

Hepatocellular carcinoma

IM:

Intrahepatic metastasis

HGF:

Hepatocyte growth factor

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Acknowledgments

We thank National Science Council in Taiwan and Research Centre of Hepatology in Buddhist Tzu Chi General Hospital for financial support.

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Authors and Affiliations

  1. Research Centre for Hepatology, Department of Internal Medicine, Buddhist Tzu Chi General Hospital and Tzu Chi University, Hualein, Taiwan

    Chi-Tan Hu, Ming-Che Lee & Siou-Mei Pan

  2. Department of Laboratory Medicine and Biotechnology, College of Medicine, Tzu Chi University, No. 701, Chung Yang Rd, Sec 3, Hualein, 970, Taiwan

    Jia-Ru Wu, Chuan-Chu Cheng, Sindy Wang, Hsiao-Ting Wang, Ling-Jung Wang, Tsu-Yao Chang & Wen-Sheng Wu

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  1. Chi-Tan Hu
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Correspondence to Wen-Sheng Wu.

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Hu, CT., Wu, JR., Cheng, CC. et al. Reactive oxygen species-mediated PKC and integrin signaling promotes tumor progression of human hepatoma HepG2. Clin Exp Metastasis 28, 851–863 (2011). https://doi.org/10.1007/s10585-011-9416-6

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  • DOI: https://doi.org/10.1007/s10585-011-9416-6

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