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Downregulation of hepatoma-derived growth factor activates the Bad-mediated apoptotic pathway in human cancer cells

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Abstract

Hepatoma-derived growth factor (HDGF) is highly expressed in human cancer and its expression is correlated with poor prognosis of cancer. The growth factor is known to stimulate cell growth while the underlying mechanism is however not clear. Transfection with HDGF cDNA stimulated while its specific antisense oligonucleotides repressed the growth of human hepatocellular carcinoma HepG2 cells. Furthermore, knock-down of HDGF by antisense oligos also induced apoptosis in HepG2 cells and in other human cancer cells, e.g. human squamous carcinoma A431 cells. HDGF knock-down was found to induce the expression of the pro-apoptotic protein Bad and also inactivate ERK and Akt, which in turn led to dephosphorylation of Bad at Ser-112, Ser-136, and activation of the intrinsic apoptotic pathway, i.e. depolarization of the mitochondrial membrane, release of mitochondrial cytochrome c, increase in the processing of caspase 9 and 3. As HDGF knock-down not only suppresses the growth but also induces apoptosis in human cancer cells, HDGF may therefore serve as a survival factor for human cancer cells and a potential target for cancer therapy.

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Acknowledgements

This work was supported by a Grant from the Shanghai-Hong Kong Anson Research Foundation, Hong Kong.

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  1. Wan Yee Tang

    Present address: Department of Environmental Health, University of Cincinnati Medical Center, Cincinnati, OH, USA

Authors and Affiliations

  1. Department of Biochemistry, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, China

    Tsun Yee Tsang, Wan Yee Tang, Wing Pui Tsang, Ngai Na Co, Siu Kai Kong & Tim Tak Kwok

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Correspondence to Tim Tak Kwok.

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Supplementary Fig. 1

Apoptosis induction in HepG2 cells after knock-down of HDGF with HDGF specific siRNA. The cells were transfected with HDGF siRNA (Hi) and scramble siRNA (S) for 24h. The antisense strand sequence for the HDGF siRNA is GUAUUUGUUGGCUGUUGAU-dTdT. (A) After transfection, the cells were allowed to grow for 72h followed by DNA fragmentation assay. (B) The cells were allowed to grow for 48h after transfection followed by annexin V/propidium iodide staining assay. (C) The expression level of HDGF after transfection with HDGF siRNA was determined by Western blot analysis. The experiments have been repeated at least 3 times with similar results. The one shown is the representative one (TIF 170 kb)

Supplementary Fig. 2

Apoptosis in HepG2 cells with knock-down of HDGF and/or Bad. The cells were transfected with HDGF antisense oligos with or without the Bad antisense oligos. After transfection, the cells were incubated for 48h followed by Annexin V binding assay. The % increase in apoptotic cells is calculated by subtracting the % of apoptotic cells in the group transfected with HDGF antisense oligos from that of the corresponding group transfected with the sense oligos. H-AS: transfection with HDGF antisense oligos only. H+B-AS: co-transfection with HDGF and Bad antisense oligos. The data are average from three independent experiments. Mean±SEM. *p<0.05, significantly different from those transfected with H-AS (TIF 141 kb)

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Tsang, T.Y., Tang, W.Y., Tsang, W.P. et al. Downregulation of hepatoma-derived growth factor activates the Bad-mediated apoptotic pathway in human cancer cells. Apoptosis 13, 1135–1147 (2008). https://doi.org/10.1007/s10495-008-0241-6

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