Skip to main content

Advertisement

Log in

Selective suppression of cervical cancer Hela cells by 2-O-β-d-glucopyranosyl-l-ascorbic acid isolated from the fruit of Lycium barbarum L.

  • Published:
Cell Biology and Toxicology Aims and scope Submit manuscript

Abstract

Lycium barbarum fruit has been used as a Chinese traditional medicine and dietary supplement for centuries. 2-O-β-d-Glucopyranosyl-l-ascorbic acid (AA-2βG), a novel stable vitamin C analog, is one of the main biologically active components of the fruit. In this report, we investigated the cytotoxic and antiproliferative effect of AA-2βG against cancer cells in vitro and identified the proteins with significantly differential expression in the cervical cancer cells (Hela) cultured in the presence of AA-2βG proteomic analysis. Our results demonstrated that the cytotoxic and antiproliferative activity of AA-2βG on cancer cell lines were in a cell type-, time-, and dose-dependent manner. Similar to vitamin C, the AA-2βG selectively induced cell death repressed the proliferation of Hela cells by the mechanism of cell apoptosis and cell cycle arrest induced by AA-2βG through a mechanism of stabilizing p53 protein. However, the biological activity of inhibition of cell proliferation in other malignant cancer cell lines or primary cells were varied, as demonstrated by either moderate inhibition or slight promotion following treatment with AA-2βG. Comparative analysis of the proteomic profiles and immunoblot analysis identified 15 proteins associated with repressing cell apoptosis and/or stimulating cell proliferation in Hela cells that were downregulated in the presence of AA-2βG or vitamin C. These data indicate that a mechanism of the AA-2βG and vitamin C mediated antitumor activity by downregulating the expression of proteins involved in cell apoptosis and proliferation and consequently inducing Hela cell apoptosis and cell cycle arrest, suggesting that AA-2βG and vitamin C may share a similar mechanism of inducing Hela cell apoptosis. These results also suggest that the L. barbarum fruit may be a potential dietary supplement and anticancer agent aimed at the prevention and treatment of cervical cancer.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Block G. Epidemiologic evidence regarding vitamin C and cancer. Am J Clin Nutr. 1991;54(6 Suppl):1310S.

    PubMed  CAS  Google Scholar 

  • Block KI, Mead MN. Vitamin C in alternative cancer treatment: historical background. Integr Cancer Ther. 2003;2:147.

    Article  PubMed  CAS  Google Scholar 

  • Cameron E, Pauling L, Leibovitz B. Ascorbic acid and cancer: a review. Cancer Res. 1979;39:663.

    PubMed  CAS  Google Scholar 

  • Candiano G, Bruschi M, Musante L, Santucci L, Ghiggeri GM, Carnemolla B, et al. Blue silver: a very sensitive colloidal Coomassie G-250 staining for proteome analysis. Electrophoresis. 2004;25:1327.

    Article  PubMed  CAS  Google Scholar 

  • Charette SJ, Landry J. The interaction of HSP27 with Daxx identifies a potential regulatory role of HSP27 in Fas-induced apoptosis. Ann NY Acad Sci. 2000;926:126.

    Article  PubMed  CAS  Google Scholar 

  • Chen Q, Espey MG, Krishna MC, Mitchell JB, Corpe CP, Buettner GR, et al. Pharmacologic ascorbic acid concentrations selectively kill cancer cells: action as a pro-drug to deliver hydrogen peroxide to tissues. Proc Natl Acad Sci USA. 2005;102:13604.

    Article  PubMed  CAS  Google Scholar 

  • Chen Q, Espey MG, Sun AY, Pooput C, Kirk KL, Krishna MC, et al. Pharmacologic doses of ascorbate act as a prooxidant and decrease growth of aggressive tumor xenografts in mice. Proc Natl Acad Sci USA. 2008;105:11105.

    Article  PubMed  CAS  Google Scholar 

  • Concannon CG, Gorman AM, Samali A. On the role of Hsp27 in regulating apoptosis. Apoptosis. 2003;8:61.

    Article  PubMed  CAS  Google Scholar 

  • Decker ED, Zhang Y, Cocklin RR, Witzmann FA, Wang M. Proteomic analysis of differential protein expression induced by ultraviolet light radiation in HeLa cells. Proteomics. 2003;3:2019.

    Article  PubMed  CAS  Google Scholar 

  • Dutcher SK. The tubulin fraternity: alpha to eta. Curr Opin Cell Biol. 2001;13:49.

    Article  PubMed  CAS  Google Scholar 

  • Esteban MA, Wang T, Qin B, Yang J, Qin D, Cai J, et al. Vitamin C enhances the generation of mouse and human induced pluripotent stem cells. Cell Stem Cell. 2010;6:71.

    Article  PubMed  CAS  Google Scholar 

  • Ford LP, Wright WE, Shay JW. A model for heterogeneous nuclear ribonucleoproteins in telomere and telomerase regulation. Oncogene. 2002;21:580.

    Article  PubMed  CAS  Google Scholar 

  • Ghosh C, Baker JA, Moysich KB, Rivera R, Brasure JR, McCann SE. Dietary intakes of selected nutrients and food groups and risk of cervical cancer. Nutr Cancer. 2008;60:331.

    Article  PubMed  CAS  Google Scholar 

  • Gorg A, Obermaier C, Boguth G, Harder A, Scheibe B, Wildgruber R, et al. The current state of two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis. 2000;21:1037.

    Article  PubMed  CAS  Google Scholar 

  • Guenal I, Risler Y, Mignotte B. Down-regulation of actin genes precedes microfilament network disruption and actin cleavage during p53-mediated apoptosis. J Cell Sci. 1997;110:489.

    PubMed  CAS  Google Scholar 

  • Hansen MD, Beckerle MC. Opposing roles of zyxin/LPP ACTA repeats and the LIM domain region in cell–cell adhesion. J Biol Chem. 2006;281:16178.

    Article  PubMed  CAS  Google Scholar 

  • Humphries JD, Wang P, Streuli C, Geiger B, Humphries MJ, Ballestrem C. Vinculin controls focal adhesion formation by direct interactions with talin and actin. J Cell Biol. 2007;179(5):1043.

    Article  PubMed  CAS  Google Scholar 

  • Kaura B, Bagga R, Patel FD. Evaluation of the pyruvate kinase isoenzyme tumor (Tu M2-PK) as a tumor marker for cervical carcinoma. J Obstet Gynaecol Res. 2004;30:193.

    Article  PubMed  CAS  Google Scholar 

  • Kim SC, Stice JP, Chen L, Jung JS, Gupta S, Wang Y, et al. Extracellular heat shock protein 60, cardiac myocytes, and apoptosis. Circ Res. 2009;105:1186.

    Article  PubMed  CAS  Google Scholar 

  • Kim J, Kim MK, Lee JK, Kim JH, Son SK, Song ES, et al. Intakes of vitamin A, C, and E, and beta-carotene are associated with risk of cervical cancer: a case–control study in Korea. Nutr Cancer. 2010;62:181.

    Article  PubMed  Google Scholar 

  • Landt S, Jeschke S, Koeninger A, Thomas A, Heusner T, Korlach S, et al. Tumor-specific correlation of tumor M2 pyruvate kinase in pre-invasive, invasive and recurrent cervical cancer. Anticancer Res. 2010;30:375.

    PubMed  CAS  Google Scholar 

  • Mackerras D, Irwig L, Simpson JM, Weisberg E, Cardona M, Webster F, et al. Randomized double-blind trial of beta-carotene and vitamin C in women with minor cervical abnormalities. Br J Cancer. 1999;79:1448.

    Article  PubMed  CAS  Google Scholar 

  • Mirvish SS. Effects of vitamins C and E on carcinogen formation and action, and relationship to human cancer. Basic Life Sci. 1986a;39:83.

    PubMed  CAS  Google Scholar 

  • Mirvish SS. Effects of vitamins C and E on N-nitroso compound formation, carcinogenesis, and cancer. Cancer. 1986b;58(8 Suppl):1842.

    Article  PubMed  CAS  Google Scholar 

  • Ohno S, Ohno Y, Suzuki N, Soma G, Inoue M. High-dose vitamin C (ascorbic acid) therapy in the treatment of patients with advanced cancer. Anticancer Res. 2009;29:809.

    PubMed  CAS  Google Scholar 

  • Padayatty SJ, Riordan HD, Hewitt SM, Katz A, Hoffer LJ, Levine M. Intravenously administered vitamin C as cancer therapy: three cases. CMAJ. 2006;174:937.

    Article  PubMed  Google Scholar 

  • Parcellier A, Schmitt E, Brunet M, Hammann A, Solary E, Garrido C. Small heat shock proteins HSP27 and alphaB-crystallin: cytoprotective and oncogenic functions. Antioxid Redox Signal. 2005;7:404.

    Article  PubMed  CAS  Google Scholar 

  • Pauling L. Effect of ascorbic acid on incidence of spontaneous mammary tumors and UV-light-induced skin tumors in mice. Am J Clin Nutr. 1991;54(6 Suppl):1252S.

    PubMed  CAS  Google Scholar 

  • Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001;29:e45.

    Article  PubMed  CAS  Google Scholar 

  • Potischman N, Brinton LA. Nutrition and cervical neoplasia. Cancer Causes Control. 1996;7:113.

    Article  PubMed  CAS  Google Scholar 

  • Ramagli L. Proteome analysis. In: Link A, editor. Methods in molecular biology, vol. 112. Totowa: Humana; 1999. p. 99.

    Google Scholar 

  • Reddy VG, Khanna N, Singh N. Vitamin C augments chemotherapeutic response of cervical carcinoma HeLa cells by stabilizing P53. Biochem Biophys Res Commun. 2001;282:409.

    Article  PubMed  CAS  Google Scholar 

  • Rivenzon-Segal D, Margalit R, Degani H. Glycolysis as a metabolic marker in orthotopic breast cancer, monitored by in vivo (13)C MRS. Am J Physiol Endocrinol Metab. 2002;283:E623.

    PubMed  CAS  Google Scholar 

  • Rock CL, Michael CW, Reynolds RK, Ruffin MT. Prevention of cervix cancer. Crit Rev Oncol Hematol. 2000;33:169.

    Article  PubMed  CAS  Google Scholar 

  • Rock CL, Moskowitz A, Huizar B, Saenz CC, Clark JT, Daly TL, et al. High vegetable and fruit diet intervention in premenopausal women with cervical intraepithelial neoplasia. J Am Diet Assoc. 2001;101:1167.

    Article  PubMed  CAS  Google Scholar 

  • Romney SL, Duttagupta C, Basu J, Palan PR, Karp S, Slagle NS, et al. Plasma vitamin C and uterine cervical dysplasia. Am J Obstet Gynecol. 1985;151:976.

    PubMed  CAS  Google Scholar 

  • Schroeder A, Mueller O, Stocker S, Salowsky R, Leiber M, Gassmann M, et al. The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol Biol. 2006;7:3.

    Article  PubMed  Google Scholar 

  • Shaw AC, Rossel Larsen M, Roepstorff P, Holm A, Christiansen G, Birkelund S. Mapping and identification of HeLa cell proteins separated by immobilized pH-gradient two-dimensional gel electrophoresis and construction of a two-dimensional polyacrylamide gel electrophoresis database. Electrophoresis. 1999;20:977.

    Article  PubMed  CAS  Google Scholar 

  • Shi Y, Zhao Y, Deng H. Powering reprogramming with vitamin C. Cell Stem Cell. 2010;6:1.

    Article  PubMed  Google Scholar 

  • Sinha P, Poland J, Kohl S, Schnolzer M, Helmbach H, Hutter G, et al. Study of the development of chemoresistance in melanoma cell lines using proteome analysis. Electrophoresis. 2003;24:2386.

    Article  PubMed  CAS  Google Scholar 

  • Takebayashi J, Yagi Y, Ishii R, Abe S, Yamada K, Tai A. Antioxidant properties of 2-O-beta-D-glucopyranosyl-L-ascorbic acid. Biosci Biotechnol Biochem. 2008;72:1558.

    Article  PubMed  CAS  Google Scholar 

  • Thomas CG, Vezyraki PE, Kalfakakou VP, Evangelou AM. Vitamin C transiently arrests cancer cell cycle progression in S phase and G2/M boundary by modulating the kinetics of activation and the subcellular localization of Cdc25C phosphatase. J Cell Physiol. 2005;205:310.

    Article  PubMed  CAS  Google Scholar 

  • Toyoda-Ono Y, Maeda M, Nakao M, Yoshimura M, Sugiura-Tomimori N, Fukami H. 2-O-(beta-D-Glucopyranosyl)ascorbic acid, a novel ascorbic acid analogue isolated from Lycium fruit. J Agric Food Chem. 2004;52:2092.

    Article  PubMed  CAS  Google Scholar 

  • Toyada-Ono Y, Maeda M, Nakao M, Yoshimura M, Sugiura-Tomimori N, Fukami H, et al. A novel vitamin C analog, 2-O-(beta-D-glucopyranosyl)ascorbic acid: examination of enzymatic synthesis and biological activity. J Biosci Bioeng. 2005;99:361.

    Article  PubMed  CAS  Google Scholar 

  • Visa N, Alzhanova-Ericsson AT, Sun X, Kiseleva E, Bjorkroth B, Wurtz T, et al. A pre-mRNA-binding protein accompanies the RNA from the gene through the nuclear pores and into polysomes. Cell. 1996;84:253.

    Article  PubMed  CAS  Google Scholar 

  • Wang Y, Zhao H, Sheng X, Gambino PE, Costell B, Bojanowski K. Protective effect of Fructus lycii polysaccharides against time and hyperthermia-induced damage in cultured seminiferous epithelium. J Ethnopharmacol. 2002;82:169.

    Article  PubMed  CAS  Google Scholar 

  • Wassertheil-Smoller S, Romney SL, Wylie-Rosett J, Slagle S, Miller G, Lucido D, et al. Dietary vitamin C and uterine cervical dysplasia. Am J Epidemiol. 1981;114:714.

    PubMed  CAS  Google Scholar 

  • Yu MS, Lai CS, Ho YS, Zee SY, So KF, Yuen WH, et al. Characterization of the effects of anti-aging medicine Fructus lycii on beta-amyloid peptide neurotoxicity. Int J Mol Med. 2007;20:261.

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China (20762007) and the Scientific and Technological Research Programs for International Cooperation of Ningxia Hui Autonomous Region.

Conflict of interest

The authors declare that there are no conflicts of interest.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yujiong Wang.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, Z., Liu, X., Wu, T. et al. Selective suppression of cervical cancer Hela cells by 2-O-β-d-glucopyranosyl-l-ascorbic acid isolated from the fruit of Lycium barbarum L.. Cell Biol Toxicol 27, 107–121 (2011). https://doi.org/10.1007/s10565-010-9174-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10565-010-9174-2

Keywords

Navigation