A modified method for purification of N-acetylgalactosamine specific lectin from Butea monosperma seeds and its effect on human hepatocellular carcinoma cell growth

Abstract

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.