Skip to main content
SpringerLink
Log in

A new Phaseolus vulgaris lectin induces selective toxicity on human liver carcinoma Hep G2 cells

  • Molecular Toxicology
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

We describe here the purification and characterization of a new Phaseolus vulgaris lectin that exhibits selective toxicity to human hepatoma Hep G2 cells and lacks significant toxicity on normal liver WRL 68 cells. This polygalacturonic acid–specific lectin (termed BTKL) was purified from seeds of P. vulgaris cv. Blue tiger king by liquid chromatography techniques. The 60-kDa dimeric lectin showed strong and broad-spectrum hemagglutinating activity toward human, rabbit, rat, and mouse erythrocytes. Bioinformatic analysis unveils substantial N-terminal sequence similarity of BTKL to other Phaseolus lectins. Among a number of tumor cells tested, BTKL exhibits potent anti-Hep G2 activity which is associated with (1) induction of DNA fragmentation, (2) production of apoptotic bodies and chromatin condensation, (3) triggering of cell apoptosis and necrosis, and (4) depolarization of mitochondrial membrane (low ΔΨm). Furthermore, BTKL could induce inducible nitric oxide synthase (iNOS) expression and subsequent nitric oxide production in vitro in mouse macrophages, which may contribute to its antitumor activity. In addition, BTKL could bring about a significant dose-dependent increase in the production of mRNAs of proinflammatory cytokines including interleukin-1 beta, interleukin-2, tumor necrosis factor alpha, and interferon-gamma. In sum, the antitumor activity and mechanism of BTKL provided here suggest that it has potential therapeutic value for human liver cancer.

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

Access this article

Log in via an institution

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

Abbreviations

BTKL:

Phaseolus vulgaris cv. Blue tiger king lectin

RT-PCR:

Reverse transcription polymerase chain reaction

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

NO:

Nitric oxide

ΔΨm:

Mitochondrial transmembrane potential

EDTA:

Ethylenediaminetetraacetic acid

References

  • Abbott DW, Hrynuik S, Boraston AB (2007) Identification and characterization of a novel periplasmic polygalacturonic acid binding protein from Yersinia enterolitica. J Mol Biol 367:1023–1033

    Article  PubMed  CAS  Google Scholar 

  • Benhamou N, Chamberland H, Pauze FJ (1990) Implication of pectic components in cell surface interactions between tomato root cells and Fusarium oxysporum f. sp. radicis-lycopersici: a cytochemical study by means of a lectin with polygalacturonic acid-binding specificity. Plant Physiol 92:995–1003

    Article  PubMed  CAS  Google Scholar 

  • Blair MW, Diaz LM, Buendia HF, Duque MC (2009) Genetic diversity, seed size associations and population structure of a core collection of common beans (Phaseolus vulgaris L.). Theor Appl Genet 119:955–972

    Article  PubMed  CAS  Google Scholar 

  • Broughton WJ, Hernandez G, Blair MW, Beebe SE, Gepts P, Vanderleyden J (2003) Beans (Phaseolus spp.)-model food legumes. Plant Soil 252:55–128

    Article  CAS  Google Scholar 

  • Burdelya L, Kujawski M, Niu G, Zhong B, Wang T, Zhang S, Kortylewski M, Shain K, Kay H, Djeu J, Dalton W, Pardoll D, Wei S, Yu H (2005) Stat3 activity in melanoma cells affects migration of immune effector cells and nitric oxide-mediated antitumor effects. J Immunol 174:3925–3931

    PubMed  CAS  Google Scholar 

  • Cheung JY, Ong RC, Suen YK, Ooi V, Wong HN, Mak TC, Fung KP, Yu B, Kong SK (2005) Polyphyllin D is a potent apoptosis inducer in drug-resistant HepG2 cells. Cancer Lett 217:203–211

    Article  PubMed  Google Scholar 

  • De Hoff PL, Brill LM, Hirsch AM (2009) Plant lectins: the ties that bind in root symbiosis and plant defense. Mol Genet Genomics 282:1–15

    Article  PubMed  Google Scholar 

  • De Mejia EG, Del Carmen Valadez-Vega M, Reynoso-Camacho R, Loarca-Pina G (2005) Tannins, trypsin inhibitors and lectin cytotoxicity in tepary (Phaseolus acutifolius) and common (Phaseolus vulgaris) beans. Plant Foods Hum Nutr 60:137–145

    Article  PubMed  CAS  Google Scholar 

  • Ercolani G, Grazi GL, Pinna AD (2010) Liver transplantation for benign hepatic tumors: a systematic review. Dig Surg 27:68–75

    Article  PubMed  Google Scholar 

  • Fang EF, Ng TB (2011) Ribonucleases of different origins with a wide spectrum of medicinal applications. Biochim Biophys Acta-Rev Cancer 1815:65–74

    CAS  Google Scholar 

  • Fang EF, Lin P, Wong JH, Tsao SW, Ng TB (2010a) A lectin with anti-HIV-1 reverse transcriptase, antitumor, and nitric oxide inducing activities from seeds of Phaseolus vulgaris cv. extralong autumn purple bean. J Agric Food Chem 58:2221–2229

    Article  PubMed  CAS  Google Scholar 

  • Fang EF, Wong JH, Bah CS, Lin P, Tsao SW, Ng TB (2010b) Bauhinia variegata var. variegata trypsin inhibitor: from isolation to potential medicinal applications. Biochem Biophys Res Commun 396:806–811

    Article  PubMed  CAS  Google Scholar 

  • Fang EF, Wong JH, Lin P, Ng TB (2010c) Biochemical and functional properties of a lectin purified from Korean large black soybeans—a cultivar of glycine max. Protein Pept Lett 17:690–698

    Article  PubMed  CAS  Google Scholar 

  • Fang EF, Wong JH, Lin P, Ng TB (2010d) Biochemical characterization of the RNA-hydrolytic activity of a pumpkin 2S albumin. FEBS Lett 584:4089–4096

    Article  PubMed  CAS  Google Scholar 

  • Frankfurt OS, Krishan A (2001) Identification of apoptotic cells by formamide-induced dna denaturation in condensed chromatin. J Histochem Cytochem 49:369–378

    Article  PubMed  CAS  Google Scholar 

  • Games PD, Dos Santos IS, Mello EO, Diz MS, Carvalho AO, de Souza-Filho GA, Da Cunha M, Vasconcelos IM, Ferreira Bdos S, Gomes VM (2008) Isolation, characterization and cloning of a cDNA encoding a new antifungal defensin from Phaseolus vulgaris L. seeds. Peptides 29:2090–2100

    Article  PubMed  CAS  Google Scholar 

  • Giannelli G, Mazzocca A, Fransvea E, Lahn M, Antonaci S (2011) Inhibiting TGF-beta signaling in hepatocellular carcinoma. Biochim Biophys Acta 1815:214–223

    PubMed  CAS  Google Scholar 

  • Hemish J, Nakaya N, Mittal V, Enikolopov G (2003) Nitric oxide activates diverse signaling pathways to regulate gene expression. J Biol Chem 278:42321–42329

    Article  PubMed  CAS  Google Scholar 

  • Jacobs P, Radzioch D, Stevenson MM (1995) Nitric oxide expression in the spleen, but not in the liver, correlates with resistance to blood-stage malaria in mice. J Immunol 155:5306–5313

    PubMed  CAS  Google Scholar 

  • Ji J, Shi J, Budhu A, Yu Z, Forgues M, Roessler S, Ambs S, Chen Y, Meltzer PS, Croce CM, Qin LX, Man K, Lo CM, Lee J, Ng IO, Fan J, Tang ZY, Sun HC, Wang XW (2009) MicroRNA expression, survival, and response to interferon in liver cancer. N Engl J Med 361:1437–1447

    Article  PubMed  CAS  Google Scholar 

  • Lam SK, Ng TB (2010a) First report of a haemagglutinin-induced apoptotic pathway in breast cancer cells. Biosci Rep 30:307–317

    Article  PubMed  CAS  Google Scholar 

  • Lam SK, Ng TB (2010b) Isolation and characterization of a French bean hemagglutinin with antitumor, antifungal, and anti-HIV-1 reverse transcriptase activities and an exceptionally high yield. Phytomedicine 17:457–462

    Article  PubMed  CAS  Google Scholar 

  • Lei HY, Chang CP (2007) Induction of autophagy by concanavalin A and its application in anti-tumor therapy. Autophagy 3:402–404

    PubMed  CAS  Google Scholar 

  • Li CQ, Wogan GN (2005) Nitric oxide as a modulator of apoptosis. Cancer Lett 226:1–15

    Article  PubMed  CAS  Google Scholar 

  • Lim S, Hung AC, Porter AG (2009) Focused PCR screen reveals p53 dependence of nitric oxide-induced apoptosis and up-regulation of maspin and plasminogen activator inhibitor-1 in tumor cells. Mol Cancer Res 7:55–66

    Article  PubMed  CAS  Google Scholar 

  • Lind DS, Kontaridis MI, Edwards PD, Josephs MD, Moldawer LL, Copeland EM III (1997) Nitric oxide contributes to adriamycin’s antitumor effect. J Surg Res 69:283–287

    Article  PubMed  CAS  Google Scholar 

  • Lioi L, Sparvoli F, Galasso I, Lanave C, Bollini R (2003) Lectin-related resistance factors against bruchids evolved through a number of duplication events. Theor Appl Genet 107:814–822

    Article  PubMed  CAS  Google Scholar 

  • Liu B, Bian HJ, Bao JK (2010) Plant lectins: potential antineoplastic drugs from bench to clinic. Cancer Lett 287:1–12

    Article  PubMed  CAS  Google Scholar 

  • Lu Z, Zhang C, Zhai Z (2005) Nucleoplasmin regulates chromatin condensation during apoptosis. Proc Natl Acad Sci USA 102:2778–2783

    Article  PubMed  CAS  Google Scholar 

  • Maestre I, Jordan J, Calvo S, Reig JA, Cena V, Soria B, Prentki M, Roche E (2003) Mitochondrial dysfunction is involved in apoptosis induced by serum withdrawal and fatty acids in the beta-cell line INS-1. Endocrinology 144:335–345

    Article  PubMed  CAS  Google Scholar 

  • Maksimovic-Ivanic D, Mijatovic S, Harhaji L, Miljkovic D, Dabideen D, Fan Cheng K, Mangano K, Malaponte G, Al-Abed Y, Libra M, Garotta G, Nicoletti F, Stosic-Grujicic S (2008) Anticancer properties of the novel nitric oxide-donating compound (S, R)-3-phenyl-4, 5-dihydro-5-isoxazole acetic acid-nitric oxide in vitro and in vivo. Mol Cancer Ther 7:510–520

    Article  PubMed  CAS  Google Scholar 

  • Narayanan S, Surendranath K, Bora N, Surolia A, Karande AA (2005) Ribosome inactivating proteins and apoptosis. FEBS Lett 579:1324–1331

    Article  PubMed  CAS  Google Scholar 

  • Neth O, Jack DL, Dodds AW, Holzel H, Klein NJ, Turner MW (2000) Mannose-binding lectin binds to a range of clinically relevant microorganisms and promotes complement deposition. Infect Immun 68:688–693

    Article  PubMed  CAS  Google Scholar 

  • Oberhammer FA, Hochegger K, Froschl G, Tiefenbacher R, Pavelka M (1994) Chromatin condensation during apoptosis is accompanied by degradation of lamin A + B, without enhanced activation of cdc2 kinase. J Cell Biol 126:827–837

    Article  PubMed  CAS  Google Scholar 

  • Okamoto M, Gohda H, Ohe G, Yoshida H, Matsuno T, Saito M, Sato M (2000) Cytokine-inducing activity and antitumor effect of a liposome-incorporated interferon-gamma-inducing molecule derived from OK-432, a streptococcal preparation. J Immunother 23:94–103

    Article  PubMed  CAS  Google Scholar 

  • Page RD (1996) TreeView: an application to display phylogenetic trees on personal computers. Comput Appl Biosci 12:357–358

    PubMed  CAS  Google Scholar 

  • Parkin DM, Bray F, Ferlay J, Pisani P (2005) Global cancer statistics, 2002. CA Cancer J Clin 55:74–108

    Article  PubMed  Google Scholar 

  • Persson A, Chang D, Crouch E (1990) Surfactant protein D is a divalent cation-dependent carbohydrate-binding protein. J Biol Chem 265:5755–5760

    PubMed  CAS  Google Scholar 

  • Pinto LS, Nagano CS, Oliveira TM, Moura TR, Sampaio AH, Debray H, Pinto VP, Dellagostin OA, Cavada BS (2008) Purification and molecular cloning of a new galactose-specific lectin from Bauhinia variegata seeds. J Biosci 33:355–363

    Article  PubMed  CAS  Google Scholar 

  • Pope RM (2002) Apoptosis as a therapeutic tool in rheumatoid arthritis. Nat Rev Immunol 2:527–535

    Article  PubMed  CAS  Google Scholar 

  • Sahara S, Aoto M, Eguchi Y, Imamoto N, Yoneda Y, Tsujimoto Y (1999) Acinus is a caspase-3-activated protein required for apoptotic chromatin condensation. Nature 401:168–173

    Article  PubMed  CAS  Google Scholar 

  • Saito S, Nakano M (1996) Nitric oxide production by peritoneal macrophages of Mycobacterium bovis BCG-infected or non-infected mice: regulatory role of T lymphocytes and cytokines. J Leukoc Biol 59:908–915

    PubMed  CAS  Google Scholar 

  • Sharon N (2008) Lectins: past, present and future. Biochem Soc Trans 36:1457–1460

    Article  PubMed  CAS  Google Scholar 

  • Sharon N, Lis H (2004) History of lectins: from hemagglutinins to biological recognition molecules. Glycobiology 14:53R–62R

    Article  PubMed  CAS  Google Scholar 

  • Suzuki T, Amano Y, Fujita M, Kobayashi Y, Dohra H, Hirai H, Murata T, Usui T, Morita T, Kawagishi H (2009) Purification, characterization, and cDNA cloning of a lectin from the mushroom Pleurocybella porrigens. Biosci Biotechnol Biochem 73:702–709

    Article  PubMed  CAS  Google Scholar 

  • Syapin PJ, Militante JD, Garrett DK, Ren L (2001) Cytokine-induced iNOS expression in C6 glial cells: transcriptional inhibition by ethanol. J Pharmacol Exp Ther 298:744–752

    PubMed  CAS  Google Scholar 

  • Tanaka M, Saijo Y, Sato G, Suzuki T, Tazawa R, Satoh K, Nukiwa T (2000) Induction of antitumor immunity by combined immunogene therapy using IL-2 and IL-12 in low antigenic Lewis lung carcinoma. Cancer Gene Ther 7:1481–1490

    Article  PubMed  CAS  Google Scholar 

  • Tanaka H, Chiba H, Inokoshi J, Kuno A, Sugai T, Takahashi A, Ito Y, Tsunoda M, Suzuki K, Takenaka A, Sekiguchi T, Umeyama H, Hirabayashi J, Omura S (2009) Mechanism by which the lectin actinohivin blocks HIV infection of target cells. Proc Natl Acad Sci USA 106:15633–15638

    Article  PubMed  CAS  Google Scholar 

  • Tarr JM, Eggleton P, Winyard PG (2006) Nitric oxide and the regulation of apoptosis in tumour cells. Curr Pharm Des 12:4445–4468

    Article  PubMed  CAS  Google Scholar 

  • Tsao SW, Wang X, Liu Y, Cheung YC, Feng H, Zheng Z, Wong N, Yuen PW, Lo AK, Wong YC, Huang DP (2002) Establishment of two immortalized nasopharyngeal epithelial cell lines using SV40 large T and HPV16E6/E7 viral oncogenes. Biochim Biophys Acta 1590:150–158

    Article  PubMed  CAS  Google Scholar 

  • Van Damme EJ, Barre A, Rouge P, Peumans WJ (2004) Cytoplasmic/nuclear plant lectins: a new story. Trends Plant Sci 9:484–489

    Article  PubMed  Google Scholar 

  • van Horssen R, Ten Hagen TL, Eggermont AM (2006) TNF-alpha in cancer treatment: molecular insights, antitumor effects, and clinical utility. Oncologist 11:397–408

    Article  PubMed  Google Scholar 

  • Verma G, Datta M (2010) IL-1beta induces ER stress in a JNK dependent manner that determines cell death in human pancreatic epithelial MIA PaCa-2 cells. Apoptosis 15:864–876

    Article  PubMed  CAS  Google Scholar 

  • Whiteman M, Rose P, Siau JL, Cheung NS, Tan GS, Halliwell B, Armstrong JS (2005) Hypochlorous acid-mediated mitochondrial dysfunction and apoptosis in human hepatoma HepG2 and human fetal liver cells: role of mitochondrial permeability transition. Free Radic Biol Med 38:1571–1584

    Article  PubMed  CAS  Google Scholar 

  • Wong JH, Ng TB (2006) Isolation and characterization of a glucose/mannose-specific lectin with stimulatory effect on nitric oxide production by macrophages from the emperor banana. Int J Biochem Cell Biol 38:234–243

    Article  PubMed  CAS  Google Scholar 

  • Wong JH, Ng TB, Cheung RC, Ye XJ, Wang HX, Lam SK, Lin P, Chan YS, Fang EF, Ngai PH, Xia LX, Ye XY, Jiang Y, Liu F (2010a) Proteins with antifungal properties and other medicinal applications from plants and mushrooms. Appl Microbiol Biotechnol 87:1221–1235

    Article  PubMed  CAS  Google Scholar 

  • Wong JH, Wan CT, Ng TB (2010b) Characterisation of a haemagglutinin from Hokkaido red bean (Phaseolus vulgaris cv. Hokkaido red bean). J Sci Food Agric 90:70–77

    Article  PubMed  CAS  Google Scholar 

  • Yang ES, Park JW (2006) Regulation of nitric oxide-induced apoptosis by sensitive to apoptosis gene protein. Free Radic Res 40:279–284

    Article  PubMed  CAS  Google Scholar 

  • Yip I, Pang XP, Berg L, Hershman JM (1995) Antitumor actions of interferon-gamma and interleukin-1 beta on human papillary thyroid carcinoma cell lines. J Clin Endocrinol Metab 80:1664–1669

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Dr. S. K. Lam for his helpful assistance in flow cytometric analysis.

Author information

Authors and Affiliations

  1. School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong

    Evandro Fei Fang, Wen Liang Pan, Jack Ho Wong, Yau Sang Chan, Xiu Juan Ye & Tzi Bun Ng

  2. Chou Ming Li Basic Medicinal Sciences Building, The Chinese University of Hong Kong, Shatin, Hong Kong

    Tzi Bun Ng

Authors
  1. Evandro Fei Fang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wen Liang Pan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jack Ho Wong
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yau Sang Chan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Xiu Juan Ye
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tzi Bun Ng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tzi Bun Ng.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 615 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fang, E.F., Pan, W.L., Wong, J.H. et al. A new Phaseolus vulgaris lectin induces selective toxicity on human liver carcinoma Hep G2 cells. Arch Toxicol 85, 1551–1563 (2011). https://doi.org/10.1007/s00204-011-0698-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-011-0698-x

Keywords

Search

Navigation