Smh-3 induces G2/M arrest and apoptosis through calcium‑mediated endoplasmic reticulum stress and mitochondrial signaling in human hepatocellular carcinoma Hep3B cells

Liu,Chin-Yu; Yang,Jai-Sing; Huang,Shih-Ming; Chiang,Jo-Hua; Chen,Ming-Hua; Huang,Li-Jiau; Ha,Ho-Yu; Fushiya,Shinji; Kuo,Sheng-Chu

doi:10.3892/or.2012.2166

Smh-3 induces G2/M arrest and apoptosis through calcium‑mediated endoplasmic reticulum stress and mitochondrial signaling in human hepatocellular carcinoma Hep3B cells

Authors:
- Chin-Yu Liu
- Jai-Sing Yang
- Shih-Ming Huang
- Jo-Hua Chiang
- Ming-Hua Chen
- Li-Jiau Huang
- Ho-Yu Ha
- Shinji Fushiya
- Sheng-Chu Kuo
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Affiliations: Graduate Institute of Pharmaceutical Chemistry, China Medical University Hospital, Taichung 404, Taiwan, R.O.C., Department of Pharmacology, China Medical University, China Medical University Hospital, Taichung 404, Taiwan, R.O.C., Department of Life Sciences, National Chung Hsing University, Taichung 402, Taiwan, R.O.C., Department of Kampo Pharmaceutical Sciences, Nihon Pharmaceutical University, Saitama 362-0806, Japan
Published online on: December 5, 2012 https://doi.org/10.3892/or.2012.2166
Pages: 751-762

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Abstract

In the present study, we investigated the antitumor effects of Smh-3 on the viability, cell cycle and apoptotic cell death in human hepatocellular carcinoma Hep3B cells in vitro. We also investigated the molecular mechanisms involved in the effects of Smh-3 on human hepatoma Hep3B cells, including the effects on protein and mRNA levels which were determined by western blotting and DNA microarray methods, respectively. The results demonstrated that Smh-3 induced growth inhibition, cell morphological changes and induction of G2/M arrest and apoptosis in Hep3B cells. DNA microarray assay identified numerous differentially expressed genes related to angiogenesis, autophagy, calcium-mediated ER stress signaling, cell adhesion, cell cycle and mitosis, cell migration, cytoskeleton organization, DNA damage and repair, mitochondrial-mediated apoptosis and cell signaling pathways. Furthermore, Smh-3 inhibited CDK1 activity, mitochondrial membrane potential (ΔΨm) and increased the cytosolic Ca2+ release and caspase-4, caspase-9 and caspase-3 activities in Hep3B cells. Western blot analysis demonstrated that Smh-3 increased the protein levels of caspase-4 and GADD153 that may lead to ER stress and consequently apoptosis in Hep3B cells. Taken together, Smh-3 acts against human hepatocellular carcinoma Hep3B cells in vitro through G2/M phase arrest and induction of calcium-mediated ER stress and mitochondrial-dependent apoptotic signaling pathways.

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