Login Register Cart 0
Generic placeholder image

Protein & Peptide Letters

Editor-in-Chief

ISSN (Print): 0929-8665
ISSN (Online): 1875-5305

Back
Journal Home About Journal Editorial Board Journal Insight Current Issue Volumes/Issues
Subscribe
Research Article

A Polylactosamine Specific Lectin from Adenia hondala Induces Apoptosis and Necrosis in Human Epithelial Colon Cancer HT-29 Cells

Author(s): Shashikala R. Inamdar*, Narasimhappagari Jagadeesh, Kavita Y. Hiremath, Shivakumar Belur and Mamta Sharma

Volume 28, Issue 10, 2021

Published on: 15 June, 2021

Page: [1108 - 1114] Pages: 7

DOI: 10.2174/0929866528666210616100140

Price: $65

Abstract

Background: Altered expression of N-glycans such as polylactosamine is observed in colon cancer. AHL, a polylactosamine specific lectin from Adenia hondala from a medicinal plant from the Passifloraceae family has been reported earlier.

Objective: The aim of the present study is to study the interaction of AHL with human colon cancer epithelial HT-29 cells and colon cancer tissues.

Methods: Cell viability was determined by MTT [3-[4, 5- dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide] assay, while cell surface binding, apoptosis by Annexin-V-PI assay and ROS production using DCFDA [2’,7’ - dichlorofluorescindiacetate] kit method were analysed by flowcytometry, immunohistochemistry was performed using biotinylated AHL, protein purification by affinity chromatography using asialofetuin-coupled Sepharose -4B column.

Results: AHL strongly binds to HT-29 cells with a Mean Fluorescence Intensity of 12.4, which could be blocked by competing glycoprotein asialofetuin. AHL inhibits HT-29 cell growth in a dose and time-dependent manner with IC50 of 2.5 μg/mL and differentially binds to human normal and cancerous tissues. AHL induces apoptosis and slight necrosis in HT-29 cells with an increase in the early apoptotic population of 25.1 and 36% for 24 h and 48 h respectively and necrotic population of 1.5 and 4.6% at 24 h and 48 h respectively as revealed by Annexin-V-PI assay. AHL induces the release of Reactive Oxygen Species in HT-29 cells in a dose-dependent manner.

Conclusion: To the best of knowledge, this is the first report on lectin from Adenia hondala which is not a RIP with apoptotic and necrotic effects. These findings support the promising potential of AHL in cancer research.

Keywords: AHL, apoptosis, necrosis, HT-29 cells, histochemistry, ROS.

« Previous Next »
Graphical Abstract

[1]
World Health Organization. GLOBOCAN 2008 Cancer incidence and mortality worldwide. International Agency for Research on Cancer. Available from: https://www.iarc.who.int/news-events/globocan-2008-cancer-incidence-and-mortality-worldwide/
[2]
World Health Organization. GLOBOCAN 2012: estimated cancer incidence, mortality and prevalence worldwide in 2012. International Agency for Research on Cancer. Available from: https://www.iarc.who.int/news-events/latest-world-cancer-statistics-globocan-2012-estimated-cancer-incidence-mortality-and-prevalence-worldwide-in-2012/
[3]
Shurer, C.R.; Colville, M.J.; Gupta, V.K.; Head, S.E.; Kai, F.; Lakins, J.N.; Paszek, M.J. Genetically encoded toolbox for glycocalyx engineering: tunable control of cell adhesion, survival, and cancer cell behaviors. ACS Biomater. Sci. Eng., 2018, 4(2), 388-399.
[http://dx.doi.org/10.1021/acsbiomaterials.7b00037] [PMID: 29805991]
[4]
Hakomori, S. Tumor-associated carbohydrate antigens. Annu. Rev. Immunol., 1984, 2(1), 103-126.
[http://dx.doi.org/10.1146/annurev.iy.02.040184.000535] [PMID: 6085749]
[5]
Feizi, T. Demonstration by monoclonal antibodies that carbohydrate structures of glycoproteins and glycolipids are onco-developmental antigens. Nature, 1985, 314(6006), 53-57.
[http://dx.doi.org/10.1038/314053a0] [PMID: 2579340]
[6]
Garrido-Abellan, F.; Duran-Ferrer, M.; MacMillan, A.; Minas, A.; Nicoletti, P.; Vecchi, G. Brucellosis in Sheep and Goats (Brucella melitensis). Scientific Committee on Animal Health and Animal Welfare – Archive. (adopted 12 July 2001). Available from: https://ec.europa.eu/food/system/files/2020-12/sci-com_scah_out59_en.pdf
[7]
Brooks, S.A. The involvement of Helix pomatia lectin (HPA) binding N-acetylgalactosamine glycans in cancer progression. Histol. Histopathol., 2000, 15(1), 143-158.
[PMID: 10668205]
[8]
Adamczyk, B.; Tharmalingam, T.; Rudd, P.M. Glycans as cancer biomarkers. Biochimica et Biophysica Acta (BBA)-. General Subjects., 2012, 1820(9), 1347-1353.
[http://dx.doi.org/10.1016/j.bbagen.2011.12.001]
[9]
Mody, R.; Joshi, S.; Chaney, W. Use of lectins as diagnostic and therapeutic tools for cancer. J. Pharmacol. Toxicol. Methods, 1995, 33(1), 1-10.
[http://dx.doi.org/10.1016/1056-8719(94)00052-6] [PMID: 7727802]
[10]
Sharma, M.; Hegde, P.; Hiremath, K.; Reddy H, V.; Kamalanathan, A.S.; Swamy, B.M.; Inamdar, S.R. Purification, characterization and fine sugar specificity of a N-Acetylgalactosamine specific lectin from Adenia hondala. Glycoconj. J., 2018, 35(6), 511-523.
[http://dx.doi.org/10.1007/s10719-018-9843-6] [PMID: 30306293]
[11]
Spiro, R.G.; Bhoyroo, V.D. Structure of the O-glycosidically linked carbohydrate units of fetuin. J. Biol. Chem., 1974, 249(18), 5704-5717.
[http://dx.doi.org/10.1016/S0021-9258(20)79875-3] [PMID: 4137945]
[12]
Goldman, M., Procedures for Conjugation In: Fluorescent Antibody Methods, 1968, 101-149.
[13]
Duk, M.; Lisowska, E.; Wu, J.H.; Wu, A.M. The biotin/avidin-mediated microtiter plate lectin assay with the use of chemically modified glycoprotein ligand. Anal. Biochem., 1994, 221(2), 266-272.
[http://dx.doi.org/10.1006/abio.1994.1410] [PMID: 7810865]
[14]
Boland, C.R.; Chen, Y.F.; Rinderle, S.J.; Resau, J.H.; Luk, G.D.; Lynch, H.T.; Goldstein, I.J. Use of the lectin from Amaranthus caudatus as a histochemical probe of proliferating colonic epithelial cells. Cancer Res., 1991, 51(2), 657-665.
[PMID: 1985783]
[15]
Matés, J.M.; Segura, J.A.; Alonso, F.J.; Márquez, J. Oxidative stress in apoptosis and cancer: An update. Arch. Toxicol., 2012, 86(11), 1649-1665.
[http://dx.doi.org/10.1007/s00204-012-0906-3] [PMID: 22811024]
[16]
Liu, B.; Li, C.Y.; Bian, H.J.; Min, M.W.; Chen, L.F.; Bao, J.K. Antiproliferative activity and apoptosis-inducing mechanism of Concanavalin A on human melanoma A375 cells. Arch. Biochem. Biophys., 2009, 482(1-2), 1-6.
[http://dx.doi.org/10.1016/j.abb.2008.12.003] [PMID: 19111670]
[17]
Yau, T.; Dan, X.; Ng, C.C.; Ng, T.B. Lectins with potential for anti-cancer therapy. Molecules, 2015, 20(3), 3791-3810.
[http://dx.doi.org/10.3390/molecules20033791] [PMID: 25730388]
[18]
Sindhura, B.R.; Hegde, P.; Chachadi, V.B.; Inamdar, S.R.; Swamy, B.M. High mannose N-glycan binding lectin from Remusatia vivipara (RVL) limits cell growth, motility and invasiveness of human breast cancer cells. Biomed. Pharmacother., 2017, 93, 654-665.
[http://dx.doi.org/10.1016/j.biopha.2017.06.081] [PMID: 28686979]
[19]
Stirpe, F.; Bolognesi, A.; Bortolotti, M.; Farini, V.; Lubelli, C.; Pelosi, E.; Polito, L.; Dozza, B.; Strocchi, P.; Chambery, A.; Parente, A.; Barbieri, L. Characterization of highly toxic type 2 ribosome-inactivating proteins from Adenia lanceolata and Adenia stenodactyla (Passifloraceae). Toxicon, 2007, 50(1), 94-105.
[http://dx.doi.org/10.1016/j.toxicon.2007.02.020] [PMID: 17434551]
[20]
Lei, H.Y.; Chang, C.P. Lectin of Concanavalin A as an anti-hepatoma therapeutic agent. J. Biomed. Sci., 2009, 16(1), 10.
[http://dx.doi.org/10.1186/1423-0127-16-10] [PMID: 19272170]
[21]
Büssing, A.; Schietzel, M. Apoptosis-inducing properties of Viscum album L. extracts from different host trees, correlate with their content of toxic mistletoe lectins. Anticancer Res., 1999, 19(1A), 23-28.
[PMID: 10226520]
[22]
Mazalovska, M.; Kouokam, J.C. Transiently expressed mistletoe lectin II in Nicotiana benthamiana demonstrates anticancer activity in vitro. Molecules, 2020, 25(11), 2562.
[http://dx.doi.org/10.3390/molecules25112562] [PMID: 32486427]
[23]
Finkel, T. Oxidant signals and oxidative stress. Curr. Opin. Cell Biol., 2003, 15(2), 247-254.
[http://dx.doi.org/10.1016/S0955-0674(03)00002-4] [PMID: 12648682]
[24]
Choi, K.; Kim, J.; Kim, G.W.; Choi, C. Oxidative stress-induced necrotic cell death via mitochondira-dependent burst of reactive oxygen species. Curr. Neurovasc. Res., 2009, 6(4), 213-222.
[http://dx.doi.org/10.2174/156720209789630375] [PMID: 19807658]
[25]
Klein, P.J.; Osmers, R.; Vierbuchen, M.; Ortmann, M.; Kania, J.; Uhlenbruck, G. The importance of lectin binding sites and carcinoembryonic antigen with regard to normal, hyperplastic, adenomatous, and carcinomatous colonic mucosa. In: Chemotherapy and Radiotherapy of Gastrointestinal Tumors. Recent Results in Cancer Research; Klein, H.O. Ed.; Springer: Berlin, Heidelberg, 1981; Vol. 79, pp.1-9
[http://dx.doi.org/10.1007/978-3-642-81681-9_1]

Rights & Permissions Print Cite
Article Metrics
15
1
© 2024 Bentham Science Publishers | Privacy Policy