Advertisement

Apigenin Down-Regulates the Hypoxia Response Genes: HIF-1α, GLUT-1, and VEGF in Human Pancreatic Cancer Cells

Published:November 29, 2010DOI:https://doi.org/10.1016/j.jss.2010.10.041

      Background

      The flavonoid apigenin exhibits anti-proliferative and anti-angiogenic activities. Our objective was to evaluate the effect of apigenin on hypoxia responsive genes important in pancreatic cancer cell proliferation.

      Materials and Methods

      Immunohistochemistry for GLUT-1 expression was conducted on human pancreatic cancer samples and adjacent controls. Real-time RT-PCR, Western blot analysis, and enzyme-linked immunosorbent assay (ELISA) were conducted on CD18 and S2-013 human pancreatic cancer cells treated with apigenin (0–50 μM) in normoxic and hypoxic conditions to evaluate HIF-1α, GLUT-1, and VEGF mRNA and protein expression and secretion.

      Results

      GLUT-1 expression was significantly increased in pancreatic adenocarcinoma samples versus adjacent controls (P < 0.001). Hypoxic conditions induced HIF-1α, GLUT-1, and VEGF protein expression in both CD18 and S2-013 pancreatic cancer cells. Apigenin (50 μM) blocked hypoxia induced up-regulation of all three proteins in both cell lines. Apigenin also impeded hypoxia-mediated induction of GLUT-1 and VEGF mRNA in both cell lines (P < 0.05).

      Conclusions

      Apigenin inhibits HIF-1α, GLUT-1, and VEGF mRNA and protein expression in pancreatic cancer cells in both normoxic and hypoxic conditions. This may account for the mechanism of apigenin’s anti-proliferative and anti-angiogenic effects and further supports the potential of apigenin as a future chemopreventive agent for pancreatic cancer.

      Key Words

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Journal of Surgical Research
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Stroszczynski C.
        • Jost D.
        • Reichardt P.
        • et al.
        Follow-up of gastro-intestinal stromal tumours (GIST) during treatment with imatinib mesylate by abdominal MRI.
        Eur Radiol. 2005; 15: 2448
        • Patel D.
        • Shukla S.
        • Gupta S.
        Apigenin and cancer chemoprevention: Progress, potential and promise.
        Int J Oncol. 2007; 30: 233
        • Melstrom L.G.
        • Salabat M.R.
        • Ding X.Z.
        • et al.
        Apigenin inhibits the GLUT-1 glucose transporter and the phosphoinositide 3-kinase/AKT pathway in human pancreatic cancer cells.
        Pancreas. 2008; 37: 426
        • Ujiki M.B.
        • Ding X.Z.
        • Salabat M.R.
        • et al.
        Apigenin inhibits pancreatic cancer cell proliferation through G2/M cell cycle arrest.
        Mol Cancer. 2006; 5: 76
        • Mirzoeva S.
        • Kim N.D.
        • Chiu K.
        • et al.
        Inhibition of HIF-1α and VEGF expression by the chemopreventive bioflavonoid apigenin is accompanied by Akt inhibition in human prostate carcinoma PC3-M cells.
        Mol Carcinog. 2008; 47: 686
        • Ren W.
        • Qiao Z.
        • Wang H.
        • et al.
        Flavonoids: Promising anticancer agents.
        Med Res Rev. 2003; 23: 519
        • Fang J.
        • Zhou Q.
        • Liu L.Z.
        • et al.
        Apigenin inhibits tumor angiogenesis through decreasing HIF-α and VEGF expression.
        Carcinogenesis. 2007; 28: 858
        • Sarkar F.H.
        • Li Y.
        Cell signaling pathways altered by natural chemopreventive agents.
        Mutat Res. 2004; 555: 53
        • Liu L.Z.
        • Fang J.
        • Zhou Q.
        • et al.
        Apigenin inhibits expression of vascular endothelial growth factor and angiogenesis in human lung cancer cells: Implication of chemoprevention of lung cancer.
        Mol Pharmacol. 2005; 68: 635
        • Osada M.
        • Imaoka S.
        • Funae Y.
        Apigenin suppresses the expression of VEGF, an important factor for angiogenesis, in endothelial cells via degradation of HIF-1α protein.
        FEBS Lett. 2004; 575: 59
        • Melillo G.
        Inhibiting hypoxia-inducible factor 1 for cancer therapy.
        Mol Cancer Res. 2006; 4: 601
      1. Talks KL, Turley H, Gatter KC, et al. The expression and distribution of the hypoxia-inducible factors HIF-1α and HIF-2α in normal human tissues, cancers, and tumor-associated macrophages. Am J Pathol 200;157:411.

        • Zhong H.
        • DeMarzo A.M.
        • Laughner E.
        • et al.
        Overexpression of hypoxia-inducible factor 1α in common human cancers and their metastases.
        Cancer Res. 1999; 59: 5830
        • Bos R.
        • Zhong H.
        • Hanrahan C.F.
        • et al.
        Levels of hypoxia-inducible factor-1α during breast carcinogenesis.
        J Natl Cancer Inst. 2001; 21: 309
        • Bos R.
        • vanderGroep P.
        • Greijer A.E.
        • et al.
        Levels of hypoxia-inducible factor-1α independently predict prognosis in patients with lymph node negative breast carcinoma.
        Cancer. 2003; 97: 1573
        • Aebersold D.M.
        • Burri P.
        • Beer K.T.
        • et al.
        Expression of hypoxia-inducible factor-1α: A novel predictive and prognostic parameter in radiotherapy of oropharyngeal cancer.
        Cancer Res. 2001; 61: 2911
        • Birner P.
        • Gatterbauer B.
        • Oberhuber G.
        • et al.
        Expression of hypoxia-inducible factor-1α in oligodendrogliomas: Its impact on prognosis and on neoangiogenesis.
        Cancer. 2001; 91: 165
        • Birner P.
        • Schindl M.
        • Obermair A.
        • et al.
        Expression of hypoxia-inducible factor 1α in epithelial ovarian tumors: Its impact on prognosis and on response to chemotherapy.
        Clin Cancer Res. 2001; 7: 1661
        • Birner P.
        • Schindl M.
        • Obermair A.
        • et al.
        Overexpression of hypoxia-inducible factor1α is a marker for an unfavorable prognosis in early-stage invasive cervical cancer.
        Cancer Res. 2000; 60: 4693
        • Koukourakis M.I.
        • Bentzen S.M.
        • Giatromanolaki A.
        • et al.
        Endogenous markers of two separate hypoxia response pathways (hypoxia inducible factor 2α and carbonic anhydrase 9) are associated with radiotherapy failure in head and neck cancer patients recruited in the CHART randomized trial.
        J Clin Oncol. 2006; 24: 727
        • Yokoi K.
        • Fidler I.J.
        Hypoxia increases resistance of human pancreatic cancer cells to apoptosis induced by gemcitabine.
        Clin Cancer Res. 2004; 10: 2299
        • Garcea G.
        • Doucas H.
        • Steward W.P.
        • et al.
        Hypoxia and Angiogenesis in Pancreatic Cancer.
        ANZ J Surg. 2006; 76: 830
        • Mabjeesh N.J.
        • Amir S.
        Hypoxia-inducible factor (HIF) in human tumorigenesis.
        Histol Histopathol. 2007; 22: 559
        • Ivan M.
        • Kondo K.
        • Yang H.
        • et al.
        HIF1α targeted for VHL-mediated destruction by proline hydroxylation: Implications for O2 sensing.
        Science. 2001; 292: 464
        • Chen C.
        • Pore N.
        • Behrooz A.
        • et al.
        Regulation of glut1 mRNA by hypoxia-inducible factor-1. Interaction between H-ras and hypoxia.
        J Biol Chem. 2001; 276: 9519
        • Karayiannakis A.J.
        • Bolanaki H.
        • Syrigos K.N.
        • et al.
        Serum vascular endothelial growth factor levels in pancreatic cancer patients correlate with advanced and metastatic disease and poor prognosis.
        Cancer Lett. 2003; 194: 119
        • Seo Y.
        • Baba H.
        • Fukuda T.
        • et al.
        High expression of vascular endothelial growth factor associated with liver metastasis and a poor prognosis for patients with ductal pancreatic adenocarcinoma.
        Cancer. 2000; 88: 2239
        • Tsuzuki Y.
        • Carreira C.M.
        • Bockhorn M.
        • et al.
        Pancreas microenvironment promotes VEGF expression and tumour growth: Novel window models for pancreatic tumour angiogenesis and microcirculation.
        Lab Invest. 2001; 81: 1439
        • Inoue M.
        • Hager J.H.
        • Ferrara N.
        • et al.
        VEGF-A has a critical, nonredundant role in angiogenic switching and pancreatic β cell carcinogenesis.
        Cancer Cell. 2002; 1: 193
        • Haber R.S.
        • Rathan A.
        • Weiser K.R.
        • et al.
        GLUT1 glucose transporter expression in colorectal carcinoma: A marker for poor prognosis.
        Cancer. 1998; 83: 34
        • Younes M.
        • Brown R.W.
        • Mody D.R.
        • et al.
        GLUT1 expression in human breast carcinoma: Correlation with known prognostic markers.
        Anticancer Res. 1995; 15: 2895
        • Kalir T.
        • Wang B.Y.
        • Goldfischer M.
        • et al.
        Immunohistochemical staining of GLUT1 in benign, borderline, and malignant ovarian epithelia.
        Cancer. 2002; 94: 1078
        • Kawamura T.
        • Kusakabe T.
        • Sugino T.
        • et al.
        Expression of glucose transporter-1 in human gastric carcinoma: Association with tumor aggressiveness, metastasis and patient survival.
        Cancer. 2001; 92: 634
        • Ito H.
        • Duxbury M.
        • Zinner M.J.
        • et al.
        Glucose transporter-1 gene expression is associated with pancreatic cancer invasiveness and MMP-2 activity.
        Surgery. 2004; 136: 548
        • Mueckler M.
        Facilitative glucose transporters.
        Eur J Biochem. 1994; 219: 713
        • Fang J.
        • Chang X.
        • Cao Z.
        • et al.
        Apigenin inhibits VEGF and HIF-1 expression via PI3K/AKT/p70S6K1 and HDM2/p53 pathways.
        FASEB J. 2005; 19: 342
        • Taniguchi S.
        • Iwamura T.
        • Katsuki T.
        Correlation between spontaneous metastatic potential and type I collagenolytic activity in a human pancreatic cancer cell line (SUIT-2) and sublines.
        Clin Exp Metastasis. 1992; 10: 259
        • Chan C.Y.
        • Salabat M.R.
        • Ding X.Z.
        Identification and in silico characterization of a novel gene: TPA-induced trans-membrane protein.
        Biochem Biophys Res Commun. 2005; 328: 755
        • Kim J.W.
        • Dang C.V.
        Cancer’s molecular sweet tooth and the Warburg Effect.
        Cancer Res. 2006; 66: 8927
        • Buchler P.
        • Reber H.A.
        • Bucher M.
        • et al.
        Hypoxia-inducible factor 1 regulates vascular endothelial growth factor expression in human pancreatic cancer.
        Pancreas. 2003; 26: 56
        • Ding X.Z.
        • Fehsenfeld D.M.
        • Murphy L.O.
        • et al.
        Physiological concentrations of insulin augment pancreatic cancer cell proliferation and glucose utilization by activating MAP kinase, PI3 kinase and enhancing GLUT-1 expression.
        Pancreas. 2000; 21: 310
        • Patiar S.
        • Harris A.L.
        Role of hypoxia-inducible factor-1a as a cancer therapy target.
        Endocr Rel Cancer. 2006; 13: S61
        • Behrooz A.
        • Ismail-Beigi F.
        Stimulation of glucose transport by hypoxia: Signals and mechanisms.
        News Physiol Sci. 1999; 14: 105
        • Asikainen T.M.
        • Ahmad A.
        • Schneider B.K.
        • et al.
        Stimulation of HIF-1a, HIF-2a, and VEGF by prolyl 4-hydroxylase inhibition in human lung endothelial and epithelial cells.
        Free Radical Biol Med. 2005; 38: 1002
        • Itakura J.
        • Ishiwata T.
        • Friess H.
        • et al.
        Enhances expression of vascular endothelial growth factor in human pancreatic cancer correlates with local disease progression.
        Clin Cancer Res. 1997; 3: 1309