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Induction of Apoptotic Effects of Antiproliferative Protein from the Seeds of Borreria hispida on Lung Cancer (A549) and Cervical Cancer (HeLa) Cell Lines

机译:诱导疏螺旋体种子抗增殖蛋白对肺癌(A549)和宫颈癌(HeLa)细胞系的凋亡作用

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摘要

A 35 KDa protein referred to as F3 was purified from the seeds of Borreria hispida by precipitation with 80% ammonium sulphate and gel filtration on Sephadex G-100 column. RP-HPLC analysis of protein fraction (F3) on an analytical C-18 column produced a single peak, detected at 220 nm. F3 showed an apparent molecular weight of 35 KDa by SDS PAGE and MALDI-TOF-MS analyses. Peptide mass fingerprinting analysis of F3 showed the closest homology with the sequence of 1-aminocyclopropane-1-carboxylate deaminase of Pyrococcus horikoshii. The protein (F3) exhibited significant cytotoxic activity against lung (A549) and cervical (HeLa) cancer cells in a dose-dependent manner at concentrations ranging from 10 µg to 1000 µg/mL, as revealed by the MTT assay. Cell cycle analysis revealed the increased growth of sub-G0 population in both cell lines exposed to a concentration of 1000 µg/mL of protein fraction F3 as examined from flow cytometry. This is the first report of a protein from the seeds of Borreria hispida with antiproliferative and apoptotic activity in lung (A549) and cervical (HeLa) cancer cells.
机译:通过用80%硫酸铵沉淀并在Sephadex G-100柱上进行凝胶过滤,从组波氏芽孢杆菌的种子中纯化出35 KDa的蛋白质,称为F3。在C-18分析柱上对蛋白质级分(F3)进行RP-HPLC分析,产生了一个单峰,在220nm处检测到。通过SDS PAGE和MALDI-TOF-MS分析,F3的表观分子量为35 KDa。 F3的肽质量指纹图谱分析显示与火球菌的1-氨基环丙烷-1-羧酸脱氨酶序列最接近。 MTT分析显示,该蛋白(F3)在浓度范围为10μg至1000μg/ mL范围内,呈剂量依赖性,对肺癌(A549)和宫颈(HeLa)癌细胞具有明显的细胞毒活性。细胞周期分析显示,流式细胞术检测到暴露于1000μg/ mL蛋白质级分F3的两种细胞系中亚G0群体的生长均增加。这是首次报道的一种来自西班牙疏螺旋体种子的蛋白质,在肺癌(A549)和宫颈癌(HeLa)癌细胞中具有抗增殖和凋亡活性。

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  • 年(卷),期 -1(2014),-1
  • 年度 -1
  • 页码 179836
  • 总页数 8
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