Chen Chiung-Ju, Shih Yung-Luen, Yeh Ming-Yang, Liao Nien-Chieh, Chung Hsueh-Yu, Liu Ko-Lin, Lee Mei-Hui, Chou Pei-Yi, Hou Hsin-Yu, Chou Jiann-Shang, Chung Jing-Gung
Jen-Teh Junior College of Medicine, Nursing and Management, Miaoli, Taiwan, R.O.C.
Department of Pathology and Laboratory Medicine, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan, R.O.C.
In Vivo. 2019 Mar-Apr;33(2):383-391. doi: 10.21873/invivo.11485.
BACKGROUND/AIM: Ursolic acid (UA), a triterpene compound present in natural plants, has been shown to induce cytotoxic effects on many human cancer cells through induction of cell-cycle arrest and apoptosis. This study investigated the effects of UA on human lung cancer NCI-H292 cells in vitro.
Flow cytometric assay was used to measure the percentage of cell viability, apoptotic cell death by double staining of annexin V and propidium iodide (PI), production of reactive oxygen species (ROS) and Ca, and mitochondriaI membrane potential (Ψ). UA-induced chromatin condensation and DNA fragmentation were examined by 4',6-diamidino-2-phenylindole staining and DNA gel electrophoresis, respectively. Western blotting was used to examine the changes of apoptosis-associated protein expression in NCI-H292 cells.
UA reduced cell viability and induced apoptotic cell death. UA increased Ca production, reduced Ψ, but did not affect ROS production in NCI-H292 cells. UA increased apoptosis-inducing factor (AIF) and endonuclease G in NCI-H292 cells.
Based on these observations, we suggest UA induces apoptotic cell death via AIF and Endo G release through a mitochondria-dependent pathway in NCI-H292 cells.
背景/目的:熊果酸(UA)是一种存在于天然植物中的三萜类化合物,已显示出通过诱导细胞周期停滞和凋亡对多种人类癌细胞产生细胞毒性作用。本研究调查了UA对人肺癌NCI-H292细胞的体外作用。
采用流式细胞术检测细胞活力百分比、通过膜联蛋白V和碘化丙啶(PI)双染检测凋亡细胞死亡、活性氧(ROS)和钙的产生以及线粒体膜电位(Ψ)。分别通过4',6-二脒基-2-苯基吲哚染色和DNA凝胶电泳检测UA诱导的染色质浓缩和DNA片段化。使用蛋白质免疫印迹法检测NCI-H292细胞中凋亡相关蛋白表达的变化。
UA降低细胞活力并诱导凋亡细胞死亡。UA增加了NCI-H292细胞中钙的产生,降低了Ψ,但不影响ROS的产生。UA增加了NCI-H292细胞中凋亡诱导因子(AIF)和核酸内切酶G的含量。
基于这些观察结果,我们认为UA通过NCI-H292细胞中线粒体依赖性途径释放AIF和Endo G诱导凋亡细胞死亡。