title: Purification and cloning of lectin that induce cell apoptosis from Allium chinense.
abstract: A 8.7 kDa lectin with high agglutin activity was isolated by affinity chromatography and cloned from Allium chinense in this study. For the MTT assay, approximately 60 µg/ml A. chinense lectin (ACL) inhibited 50% of the human hepatoma Hep-3B cells grown after 48 h. In addition, no antiproliferative effect was observed on normal human umbilical vein endothelial cells (HUVEC) even at 100 µg/ml concentration. After treatments with ACL on Hep-3B cells, morphologic changes in the nucleus and cytoskeleton were observed under laser scanning confocal microscopy with 4',6-diamidino-2-phenylindole and tubulin Alexa Fluor 488 staining; whereas, the mitochondrial membrane potential was observed through Mito Tracker Red CMXRos staining. The results showed that ACL led to cell morphology and structure change (e.g., round cell shrinkage). Moreover, ACL resulted in significant change in the shape of the nucleus, damaged the cytoskeleton when tubulin was degraded, and reduced the mitochondrial transmembrane potential. By contrast, no changes were observed on HUVEC cells under the same treatment conditions. DNA fragmentation analysis was used to detect DNA damage. Western blot showed that ACL upregulated caspase-3 and Bax expression during apoptosis and cloned the structural gene of ACL with an open reading frame of 456 bp encoding 151 amino acid residues. The results showed that ACL is a potential anticancer drug.
authors.affiliation: College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha 410081, China.
authors.forename: Xiuqing
authors.initials: X
authors.lastname: Xiao
authors.affiliation: College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha 410081, China.
authors.forename: Hao
authors.initials: H
authors.lastname: He
authors.affiliation: College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha 410081, China. Electronic address: dingxuezhi@hunnu.edu.cn.
authors.forename: Xuezhi
authors.initials: X
authors.lastname: Ding
authors.affiliation: College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha 410081, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Yang
authors.affiliation: College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha 410081, China.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Liu
authors.affiliation: College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha 410081, China.
authors.forename: Shuang
authors.initials: S
authors.lastname: Liu
authors.affiliation: College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha 410081, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Rang
authors.affiliation: College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha 410081, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.affiliation: College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha 410081, China.
authors.forename: Mingxing
authors.initials: M
authors.lastname: Zuo
authors.affiliation: College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha 410081, China. Electronic address: Xialiqiu@hunnu.edu.cn.
authors.forename: Liqiu
authors.initials: L
authors.lastname: Xia
mesh_terms: D000490:Allium / Q000737:chemistry*; D000595:Amino Acid Sequence; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D001483:Base Sequence; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000648:ultrastructure; D003001:Cloning, Molecular; D003599:Cytoskeleton / Q000648:ultrastructure; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000648:ultrastructure; D006801:Humans; D037102:Lectins / Q000494:pharmacology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000494:pharmacology; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D000972:Antineoplastic Agents, Phytogenic; C494826:BAX protein, human; D037102:Lectins; D011994:Recombinant Proteins; D051028:bcl-2-Associated X Protein; C505407:CASP3 protein, human; D053148:Caspase 3
keywords: Allium chinense; Anticancer activity; Apoptosis; Lectin