A modified method for purification of N-acetylgalactosamine specific lectin from Butea monosperma seeds and its effect on human hepatocellular carcinoma cell growth
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Butea monosperma is a medicinal plant extensively used in ayurveda and is known for its medicinal applications. Butea monosperma seed powder is used for treating various health ailments including cancer. Although, most parts of the plant are being used in medicine none of them except for seeds are known to contain lectins. A lectin from seeds has been reported earlier, however it was not tested for its anticancer properties. Since plant lectins are known for their anticancer properties, in order to explore anticancer potential of B. monosperma a legume lectin was purified from seeds in a single step by nine fold, by a modified method using affinity chromatography on lactose-Sepharose 4B column and tested for its anticancer properties against human hepatocellular carcinoma cells. SDS PAGE of BML reveals two bands with Mr of 32 and 34 kDa and a minor band of 67 kDa similar to earlier reported BML. Hapten inhibition assay shows that lectin is specific to GalNAc, galactose, lactose and D-Fucose. BML shows strong binding to Hep G2 cells with MFI of 476. MTT assay showed growth inhibitory effect on Hep G2 cells by BML, PTL1 and PTL2 in a dose and time dependent manner. Growth inhibitory effect of BML on Hep G2 cells was compared with two legume lectins, PTL1 and PTL2 from Psophocarpus tetragonolobus of Fabaceae Family. BML, PTL1 and PTL2 showed growth inhibitory effect on Hep G2 cells. The presence of lectin, in B. monosperma seeds contribute for its anticancer properties.
KeywordsButea monosperma lectin (BML) Psophocarpus tetragonolobus lectin 1 (PTL1) Psophocarpus tetragonolobus lectin 2 (PTL2) Hep G2 cells Modified method
Minimum concentration of protein required for agglutination
Minimum inhibitory concentration
Mean fluorescence intensity
SRI would like to thank UGC for the funding from under UPE (F.No14-4/2012(NS/PE) and KYH would like to thank Rajiv Gandhi National Fellowship for Students with Disabilities (RGNFD) under UGC (No. F./2015-16/R GNF-2015-17-KAR -1523).
Compliance with ethical standards
Conflict of interest
Authors don’t have any conflict of interest to declare concerning to the present work.
- Goldman M (1968) Fluroscent antibody methods. Academic Press, Nw YorkGoogle Scholar
- Lis H, Sharon N (1986) Lectins as molecules and as tools. Annu Rev Biochem 55:35–67. https://doi.org/10.1146/annurev.bi.55.070186.000343 CrossRefPubMedGoogle Scholar
- Lowry OH, Rosebrough NJ, Farr LA, Randall RJ (1951) Protein measurement with the folin phenol reagent. J BiolChem 193:265–275Google Scholar
- Mathan GF, Saxena AK, Chandan BK, Jaggi BS, Gupta BD, Qazi GN, Balasundaram C, Anand Rajan KD, Kumar VL, Kumar V (2011) Chemoprevention with aqueous extract of Butea monosperma flowers results in normalization of nuclear morphometry and inhibition of a proliferation marker in liver tumors. Phytother Res. https://doi.org/10.1002/ptr.3249 CrossRefPubMedGoogle Scholar
- Mathan G, Polachi N, Nagaraj N, Boopathi S (2015) Antiproliferative activity of n-butanol floral extract from Butea monosperma against HCT 116 colon cancer cells; drug likeness properties and in silico evaluation of their active compounds toward glycogen synthase kinase-3β/axin and β-catenin/t-cell facto 8(1). ISSN 0974-2441Google Scholar
- Mehta B, Bokadia M (1981) Monospermin: a new alkaloid of Butea monosperma seeds. Chem Ind 3:98Google Scholar
- Muzumdar PM, Das MK, Das S (2011) Butea Monosperma (Lam) Kuntz—a comprehensive review. Int J Pharm Sci Nanotechnol 4(2)Google Scholar
- Pandey PK, Singh D, Singh S, Khan MY, Jamal F (2014) A nonhost peptidase inhibitor of ~ 14 kDa from Butea monosperma (Lam.) Taub seeds affects negatively the growth and developmental physiology of Helicoverpa armigera. Biochem Res Int. https://doi.org/10.1155/2014/361821 CrossRefPubMedPubMedCentralGoogle Scholar
- Razdak MK, Kapila K, Bhinde NK (1969) Antifertility effect and some pharmacological actions of Butea frondosa seed extracts. Indian J Physiol Pharmacol 13(4):239–249Google Scholar
- Tenzin C, Shukla SK, Kumar V (2010) Chemopreventive and anticancer properties of the aqueous extract of flowers of Butea monosperma. J Ethanopharmacol 219:208–213Google Scholar