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诱导博雷里亚 Hispida 种子来源的增殖抑制蛋白对肺癌(A549)和宫颈癌(HeLa)细胞系的凋亡作用。

Induction of apoptotic effects of antiproliferative protein from the seeds of Borreria hispida on lung cancer (A549) and cervical cancer (HeLa) cell lines.

机构信息

Department of Biotechnology, School of Bioengineering, SRM University, Kattankulathur 603203, India.

出版信息

Biomed Res Int. 2014;2014:179836. doi: 10.1155/2014/179836. Epub 2014 Jan 30.

Abstract

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.

摘要

一种 35 kDa 的蛋白质 F3 被从 Borreria hispida 的种子中提取出来,方法是用 80%的硫酸铵沉淀和 Sephadex G-100 柱凝胶过滤。对蛋白质部分(F3)在分析性 C-18 柱上进行 RP-HPLC 分析,产生一个单一的峰,在 220nm 处检测到。SDS PAGE 和 MALDI-TOF-MS 分析表明,F3 的表观分子量为 35 kDa。F3 的肽质量指纹图谱分析显示与 Pyrococcus horikoshii 的 1-氨基环丙烷-1-羧酸脱氨酶序列具有最接近的同源性。该蛋白(F3)在浓度为 10μg 至 1000μg/mL 时,对肺(A549)和宫颈(HeLa)癌细胞表现出显著的细胞毒性活性,这一点通过 MTT 测定得到证实。细胞周期分析显示,在暴露于 1000μg/mL 蛋白部分 F3 的两种细胞系中,亚 G0 群体的生长增加,这是从流式细胞术检查中发现的。这是首次报道来自 Borreria hispida 种子的一种具有抗增殖和促凋亡活性的蛋白质,可作用于肺(A549)和宫颈(HeLa)癌细胞。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c680/3925513/8944a0e0ed29/BMRI2014-179836.001.jpg

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