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Induction of Apoptotic Effects of Antiproliferative Protein from the Seeds ofBorreria hispidaon Lung Cancer (A549) and Cervical Cancer (HeLa) Cell Lines

机译:抗增殖蛋白的诱导抗增殖蛋白的凋亡效应(A549)和宫颈癌(HELA)细胞系

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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%硫酸铵和凝胶过滤在Sephadex G-100柱上,从博尔雷里亚·哌啶的种子中纯化35kDa蛋白。在分析C-18柱上的蛋白质级分(F3)的RP-HPLC分析在220nm处检测到单一峰。 F3通过SDS PAGE和MALDI-TOF-MS分析显示出35kDa的表观分子量。 F3的肽质量指纹分析显示了与Pyrococcus Horikoshii的1-氨基环丙烷-1-羧酸盐序列的最接近的同源性。蛋白质(F3)在浓度为10μg至1000μg/ ml的浓度下,蛋白质(F3)对肺(A549)和宫颈(HELA)癌细胞的显着细胞毒性活性表现出对肺(A549)和宫颈(HELA)癌细胞,如MTT测定所揭示的那样。细胞循环分析显示,通过从流式细胞术中检查的,在暴露于1000μg/ ml蛋白质级分F3的浓度为1000μg/ ml蛋白质级分f3的浓度下,亚g0种群的增长增加。这是来自猪(A549)和宫颈(HELA)癌细胞的抗增殖和凋亡活性的博尔切氏植物种子的蛋白质的第一报告。

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