Park Hyun-Ji, Park Shin-Hyung
Departments of Pathology, College of Korean Medicine, Dong-eui University, Busan, Korea.
J Pharmacopuncture. 2018 Dec;21(4):268-276. doi: 10.3831/KPI.2018.21.030. Epub 2018 Dec 31.
The purpose of this study is to investigate the anti-cancer effects of different fractions of (AM) in human non-small cell lung cancer (NSCLC) cells.
We isolated hexane, ethyl acetate, and butanol fractions from crude ethanol extract of AM. The cell death was examined by MTT assay and trypan blue exclusion assay. Apoptosis was detected by DAPI staining, annexin V-PI double staining and cell cycle analysis. The expression of apoptosis-related proteins and mitogen-activated protein kinases (MAPKs) was examined by western blot.
Among various fractions of AM, the ethyl acetate fraction of AM (EAM) showed the strongest cytotoxic effect in NSCLC cells. EAM reduced the cell proliferation in a time- and dose-dependent manner in NSCLC cells. In addition, EAM induced the chromatin condensation, and increased the population of sub-G1 phase and annexin V-positive cells in a time-dependent manner, indicating that EAM induced apoptosis in NSCLC cells. Consistently, EAM enhanced the expression of cleaved caspase-8 and -9, and induced the accumulation of cleaved- poly (ADP-ribose) polymerase (PARP). Among MAPK proteins, only ERK was dephosphorylated by EAM, suggesting that ERK might be related with EAM-induced apoptosis.
Our results clearly demonstrate that EAM exhibited anti-cancer effects in NSCLC cells by induction of apoptosis. We provide a valuable evidence which suggests that AM could be a desirable therapeutic option for treatment of NSCLC.
本研究旨在探讨(AM)不同组分对人非小细胞肺癌(NSCLC)细胞的抗癌作用。
我们从AM的粗乙醇提取物中分离出正己烷、乙酸乙酯和丁醇组分。通过MTT法和台盼蓝排斥试验检测细胞死亡情况。通过DAPI染色、膜联蛋白V-碘化丙啶双染色和细胞周期分析检测细胞凋亡。通过蛋白质印迹法检测凋亡相关蛋白和丝裂原活化蛋白激酶(MAPK)的表达。
在AM的各个组分中,AM的乙酸乙酯组分(EAM)在NSCLC细胞中显示出最强的细胞毒性作用。EAM以时间和剂量依赖性方式降低NSCLC细胞的增殖。此外,EAM诱导染色质浓缩,并以时间依赖性方式增加亚G1期和膜联蛋白V阳性细胞的比例,表明EAM诱导NSCLC细胞凋亡。一致地,EAM增强了裂解的半胱天冬酶-8和-9的表达,并诱导了裂解的聚(ADP-核糖)聚合酶(PARP)的积累。在MAPK蛋白中,只有ERK被EAM去磷酸化,表明ERK可能与EAM诱导的凋亡有关。
我们的结果清楚地表明,EAM通过诱导凋亡在NSCLC细胞中表现出抗癌作用。我们提供了有价值的证据,表明AM可能是治疗NSCLC的理想治疗选择。