An alkaloid
(–)-Securinine is an alkaloid originally isolated from S. suffructicosa.1 It reduces proliferation of SW480 colon adenocarcinoma cells in a dose- and time-dependent manner via increased Beclin 1 expression and induction of autophagy.2 It decreases viability of HL-60 leukemia cells (IC50s = 47.88, 23.85, and 18.87 μM at 24, 48, and 72 hours post-treatment, respectively).3 It also induces cell cycle arrest at the G1/S phase and decreases PI3K, Akt, and mTOR gene expression in a dose-dependent manner. (–)-Securinine inhibits GABA and GABA-activated benzodiazepine binding to rat brain membranes (IC50s = 57 and 101 μM, respectively).1 In vivo, (–)-securinine (5.2 mg/kg) induces clonic and tonic convulsions in mice, an effect that is reversed by administration of GABA.4
1.Beutler, J.A., Karbon, E.W., Brubaker, A.N., et al.Securinine alkaloids: A new class of GABA receptor antagonistBrain Res.330(1)135-140(1985) 2.Xia, Y.-H., Chang, C.-R., Yao, S.-Y., et al.L-securinine induced the human colon cancer SW480 cell autophagy and its molecular mechanismFitoterapia82(8)1258-1264(2011) 3.Han, S., Zhang, G., Li, M., et al.L-securinine induces apoptosis in the human promyelocytic leukemia cell line HL-60 and influences the expression of genes involved in the PI3K/AKT/mTOR signaling pathwayOncol. Rep.31(5)2245-2251(2014) 4.Tao, S.-C., Peng, J.-Z., and Lu, M.-W.Central convulsive action of l-securinineZhongguo Yao Li Xue Bao.7(1)9-12(1986)
Kinase experiment: | The cells are seeded in 12-well plates (1×105/well) and treated with (-)-Securinine at concentrations of 1.0 to 50.0 μg/mL. The control cells are exposed to DMSO at a concentration of 0.5% (v/v). After 6 h and 24 h of exposure, the activity of caspase-9 is measured by Caspase-Glo 9 Assay Kit and Glomax Multi+ Detection System, according to the manufacturer’s instruction. The activity of caspase-3/7 is assessed after 24 h of exposure the cells to (-)-Securinine. Then the cells are harvested and prepared using Muse Caspase-3/7 Assay Kit according with the manufacturer’s protocol. The stained cells are analyzed by Muse Cell Analyzer. The experiments are performed at least in three independent repeats[1]. |
Cell experiment: | The viability of the cells is determined by MTT assay. HeLa cells are seeded in 96-well plates at a density of 5×103 cells/well and treated for 24 h with (-)-Securinine in the concentration range of 1.0 to 20.0 μg/mL. The maximal concentrations of the solvents used in all the MTT experiments are 5.0% (v/v) and 1.0% (v/v) for methanol and DMSO, respectively. The absorption of the obtained formazan solution is measured with a plate reader. The viability results are presented as IC50 mean values of at least three independent experiments[1]. |
Animal experiment: | 6 week old female nude mice are used and injected bilaterally s.c. with 10×106 HL-60 cells. (-)-Securinine treatment is started 10 days after tumor cell injection. Palpable tumors are present for the established tumor model prior to initiating drug treatment. 15 mg/kg of (-)-Securinine or vehicle (30 μL of DMSO and 70 μL of water) are injected i.p. 2 or 3 times a day for 5 days followed by once a day for two days. This injection schedule is repeated for two additional weeks[2]. |
参考文献: [1]. Stefanowicz-Hajduk J, et al. Securinine from Phyllanthus glaucus Induces Cell Cycle Arrest and Apoptosis in Human Cervical Cancer HeLa Cells. PLoS One. 2016 Oct 28;11(10):e0165372. | |
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