Lectin from green speckled lentil seeds (Lens culinaris) triggered apoptosis in nasopharyngeal carcinoma cell lines

Abstract

BackgroundThe green speckled lentil seed (Lens culinaris) lectin (GSLL) exhibits hemagglutinating activity, and possesses some properties distinct from those of other lentil lectins (e.g., molecular size, biological activities) that deserve further investigation. This study aims to investigate the basic properties (e.g., molecular size, amino acid sequence, sugar specificity) and biological activities (e.g., antiproliferative activity) of GSLL.MethodsGSLL was purified by successive fractionation on SP-Sepharose, Affi-gel blue gel, Mono Q, and Superdex 75. The biochemical properties of GSLL were investigated by SDS-PAGE, mass spectrometry, N-terminal amino acid sequencing, and sugar inhibition tests. For the biological activities, purified lyophilized GSLL was sterilized, adjusted to concentrations from 1 to 0 mg/mL (by twofold serial dilution) in Dulbecco’s modified Eagle’s medium with fetal bovine serum, and examined by using the MTT assay, flow cytometry, and western blotting after treatment of nasopharyngeal carcinoma CNE1 and CNE2 cell lines with the lectin.ResultsGSLL appeared as a 21-kDa band in non-reducing SDS-PAGE. It was composed of two subunits with molecular sizes of 17 and ~4 kDa. It exhibited specificity in binding to glucose and mannose, as well as glucosides and mannosides. Mass spectrometry and N-terminal amino acid sequencing revealed similarity of GSLL to L. culinaris lectin (LcL), especially higher coverage of the β-chain of LcL. A 48-h treatment with GSLL exerted antiproliferative effects on nasopharyngeal carcinoma CNE1 and CNE2 cell lines with significant inhibition at 0.125 mg/mL (P < 0.001) and 1 mg/mL (P = 0.004), respectively, and these effects were attenuated in the presence of glucose and mannose. GSLL induced apoptosis in nasopharyngeal carcinoma CNE1 cells, with detectable phosphatidylserine externalization, mitochondrial depolarization, and cell cycle arrest. Western blot analysis suggested that GSLL triggered the extrinsic apoptotic pathway involving caspase 3, 8, and 9 in CNE1 cells.ConclusionGSLL possessed some different properties from LcL (e.g., lower pI), and increased caspase 3, 8, and 9 activity in CNE1 cells.

Pea, Pisum sativum, and its anticancer activity

“…In several review articles have mentioned the health benefits of P. sativum due to its concentration and properties of starch, protein, fiber, vitamins, minerals, and phytochemicals. [ 2 44 ] In addition, the plants in the same Fabaceae family also show the anticancer activities such as alfalfa, Medicago sativa ;[ 45 ] carob, Ceratonia siliqua ;[ 46 ] lentil, Lens culinaris ;[ 47 ] and soybean, Glycine max . [ 48 ]…”

DPP-IV Inhibitory Potentials of Flavonol Glycosides Isolated from the Seeds of Lens culinaris: In Vitro and Molecular Docking Analyses

“…Lentil has long been cultivated and used as foodstuff in Europe, the Middle East, Africa, and Asia [ 9 ]. Lentil was reported to have diverse biological activities such as antioxidant [ 9 , 10 ], α-glucosidase-inhibitory [ 10 ], anti-inflammatory [ 11 ], and anticancer effects [ 12 ]. In a previous phytochemical study, phenolics, flavonoids, tannins, saponins, and fatty acids were isolated from L. culinaris [ 9 , 13 , 14 , 15 ].…”

Inhibitory Effects of Pulse Bioactive Compounds on Cancer Development Pathways

“…The pathway involved upregulation of caspase 3, 8 and 9 in cells affected by WKBL treatment. Lectin found in lentils has demonstrated apoptotic activity through the extrinsic apoptotic pathway, the mechanism of action was through caspase 3, 8, and 9 as detected in CNE1 cells [ 70 ]. Additional apoptotic pathways were identified by phosphatidylserine externalization, mitochondrial depolarization and cell cycle arrest [ 70 ].…”