Department of Biotechnology, Alagappa University, Karaikudi 630 003, Tamil Nadu, India.
Department of Biotechnology, Alagappa University, Karaikudi 630 003, Tamil Nadu, India.
Biomed Pharmacother. 2018 Sep;105:742-752. doi: 10.1016/j.biopha.2018.06.035. Epub 2018 Jun 13.
In the present study, the antiproliferative activity of phytol and its mechanism of action against human lung adenocarcinoma cell line A549 were studied in detail. Results showed that phytol exhibited potent antiproliferative activity against A549 cells in a dose and time-dependent manner with an IC value of 70.81 ± 0.32 μM and 60.7 ± 0.47 μM at 24 and 48 h, respectively. Phytol showed no adverse toxic effect in normal human lung cells (L-132), but mild toxic effect was observed when treated with maximum dose (67 and 84 μM). No membrane-damaging effect was evidenced by PI staining and SEM analysis. The results of mitochondrial membrane potential analysis, cell cycle analysis, FT-IR and Western blotting analysis clearly demonstrated the molecular mechanism of phytol as induction of apoptosis in A549 cells, as evidenced by formation of shrinked cell morphology with membrane blebbing, depolarization of mitochondrial membrane potential, increased cell population in the sub-G0 phase, band variation in the DNA and lipid region, downregulation of Bcl-2, upregulation of Bax and the activation of caspase-9 and -3. In addition, phytol inhibited the CAM vascular growth as evidenced by CAM assay, which positively suggests that phytol has anti-angiogenic potential. Taken together, these findings clearly demonstrate the mode of action by which phytol induces cell death in A549 lung adenocarcinoma cells.
在本研究中，详细研究了叶绿醇对人肺腺癌细胞系 A549 的增殖抑制活性及其作用机制。结果表明，叶绿醇对 A549 细胞表现出强烈的增殖抑制活性，呈剂量和时间依赖性，在 24 和 48 h 时的 IC 值分别为 70.81±0.32 μM 和 60.7±0.47 μM。叶绿醇对正常人体肺细胞（L-132）无不良毒性作用，但在最大剂量（67 和 84 μM）下观察到轻微毒性作用。PI 染色和 SEM 分析未显示出膜损伤作用。线粒体膜电位分析、细胞周期分析、FT-IR 和 Western blot 分析的结果清楚地表明了叶绿醇作为 A549 细胞凋亡诱导物的分子机制，表现为细胞形态皱缩、膜起泡、线粒体膜电位去极化、亚 G0 期细胞群体增加、DNA 和脂质区带变化、Bcl-2 下调、Bax 上调和 caspase-9 和 caspase-3 激活。此外，叶绿醇通过 CAM 测定抑制 CAM 血管生长，这表明叶绿醇具有抗血管生成潜力。综上所述，这些发现清楚地表明了叶绿醇诱导 A549 肺腺癌细胞死亡的作用模式。
