title: Anti-proliferative and apoptotic effects of the novel taspine derivative tas41 in the Caco-2 cell line.
abstract: Taspine was screened and isolated for the first time from Radix et Rhizoma Leonticis. Tas41 is a novel taspine derivative. We investigated the effects of tas41 on proliferation of the Caco-2 cell line using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), a fluorescence-activated cell sorter (FACS), enzyme-linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR) and western blotting (WB). Changes in the cell cycle, apoptosis, activation of caspase-3, caspase-8 and caspase-9, and expressions of vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) were investigated after Caco-2 cells were treated with tas41. At the same time, expressions of apoptosis protein bcl-2 and bax were determined. Tas41 was found to induce apoptosis in a concentration-dependent manner as confirmed by DNA fragmentation analysis, TUNEL assay and flow cytometry. Protein and mRNA expressions of EGF, VEGF, CDK2, bcl-2 and bax were evaluated by ELISA, WB and RT-PCR. Tas41 had a better anti-proliferative effect than taspine on Caco-2 cells. A DNA ladder and apoptosis was observed, and the increased apoptotic activity by tas41 was accompanied by a decrease in the expression of VEGF protein and mRNA. The activities of caspase-3, caspase-8 and caspase-9 were significantly increased in cells treated with tas41 compared with those in the control group. In addition, protein and mRNA expressions of bcl-2 were decreased, and protein and mRNA expressions of bax were increased. These findings demonstrate that tas41 can inhibit the proliferation of, and induce apoptosis in, Caco-2 cells by activating caspase-3, caspase-8 and caspase-9, downregulating the expressions of VEGF, upregulating the ratio of bax/bcl-2.
authors.affiliation: School of Medicine, Xi'an Jiaotong University, No. 76, Yanta Weststreet, #120, Xi'an, Shaanxi Province 710061, PR China.
authors.forename: Yanmin
authors.initials: Y
authors.lastname: Zhang
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Bingling
authors.initials: B
authors.lastname: Dai
authors.forename: Nan
authors.initials: N
authors.lastname: Wang
authors.forename: Langchong
authors.initials: L
authors.lastname: He
mesh_terms: D000470:Alkaloids / Q000737:chemistry / Q000494:pharmacology*; D000972:Antineoplastic Agents, Phytogenic / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D018938:Caco-2 Cells; D053148:Caspase 3 / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D019938:Cyclin D1 / Q000096:biosynthesis; D053938:DNA Fragmentation; D015536:Down-Regulation / Q000187:drug effects; D005434:Flow Cytometry; D006801:Humans; D020287:In Situ Nick-End Labeling; D009025:Morpholines / Q000737:chemistry / Q000494:pharmacology*; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation / Q000187:drug effects; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis / Q000235:genetics; D051028:bcl-2-Associated X Protein / Q000096:biosynthesis
chemical_list: D000470:Alkaloids; D000972:Antineoplastic Agents, Phytogenic; D009025:Morpholines; D042461:Vascular Endothelial Growth Factor A; D051028:bcl-2-Associated X Protein; C569588:tas41 compound; D019938:Cyclin D1; D012313:RNA; D053148:Caspase 3; D053181:Caspase 8; D053453:Caspase 9; C018394:taspine