Wu Tsu-Tuan, Lu Chien-Lin, Lin Hen-I, Chen Bing-Fang, Jow Guey-Mei
Department of Respiratory Therapy, Fu Jen Catholic University, New Taipei City, Taiwan, R.O.C.
Division of Nephrology, Department of Medicine, Shin‑Kong Wu Ho‑Su Memorial Hospital, Taipei, Taiwan, R.O.C.
Oncol Rep. 2016 Jan;35(1):227-34. doi: 10.3892/or.2015.4368. Epub 2015 Oct 30.
β-elemonic acid, a known triterpene, exhibits anti-inflammatory effects, yet research on the pharmacological effects of β-elemonic acid is rare. We investigated the anticancer effects and the related molecular mechanisms of β-elemonic acid on human non-small cell lung cancer (NSCLC) A549 cells. The effects of β-elemonic acid on the growth of A549 cells were studied using a 3-(4,5)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was detected using Annexin V staining. The effect of β-elemonic acid on the cell cycle of A549 cells was assessed using the propidium iodide method. The change in reactive oxygen species (ROS) was detected using a dichlorodihydrofluorescein diacetate (DCFH-DA) assay with microscopic examination. The expression levels of Bcl-2 family proteins, mitogen-activated protein kinase (MAPK) family proteins and cyclooxygenase 2 (COX-2) were detected using western blot analysis. Our data revealed that β-elemonic acid strongly induced human A549 lung cancer cell death in a dose- and time-dependent manner as determined by the MTT assay. β-elemonic acid-induced cell death was considered to be apoptotic when the phosphatidylserine exposure was observed using Annexin V staining. The death of human A549 lung cancer cells was caused by apoptosis induced by activation of ROS activity, increase in the sub-G1 proportion, downregulation of Bcl-2 expression, upregulation of Bax expression and inhibition of the MAPK signaling pathways. These results clearly demonstrated that β-elemonic acid inhibits proliferation by inducing hypoploid cells and cell apoptosis. Moreover, the anticancer effects of β-elemonic acid were related to the MAPK signaling pathway, ROS activation and glutathione depletion in human A549 lung cancer cells.
β-榄香烯酸是一种已知的三萜类化合物，具有抗炎作用，但关于β-榄香烯酸药理作用的研究却很少。我们研究了β-榄香烯酸对人非小细胞肺癌（NSCLC）A549细胞的抗癌作用及其相关分子机制。采用3-(4,5)-2,5-二苯基四氮唑溴盐（MTT）法研究β-榄香烯酸对A549细胞生长的影响。使用膜联蛋白V染色检测细胞凋亡。采用碘化丙啶法评估β-榄香烯酸对A549细胞周期的影响。使用二氯二氢荧光素二乙酸酯（DCFH-DA）法并通过显微镜检查检测活性氧（ROS）的变化。采用蛋白质免疫印迹分析检测Bcl-2家族蛋白、丝裂原活化蛋白激酶（MAPK）家族蛋白和环氧化酶2（COX-2）的表达水平。我们的数据显示，MTT法测定结果表明，β-榄香烯酸以剂量和时间依赖性方式强烈诱导人A549肺癌细胞死亡。当使用膜联蛋白V染色观察到磷脂酰丝氨酸暴露时，β-榄香烯酸诱导的细胞死亡被认为是凋亡性的。人A549肺癌细胞的死亡是由ROS活性激活、亚G1期比例增加、Bcl-2表达下调、Bax表达上调以及MAPK信号通路受抑制诱导的凋亡所致。这些结果清楚地表明，β-榄香烯酸通过诱导亚倍体细胞和细胞凋亡来抑制增殖。此外，β-榄香烯酸的抗癌作用与人A549肺癌细胞中的MAPK信号通路、ROS激活和谷胱甘肽耗竭有关。
