Wu Zi-Li, Yin Zhong-Qiong, Du Yong-Hua, Feng Rui-Zhang, Ye Kui-Chuan, Wei Qin, Hu Yong, He Lin, Liao Lin, Wang Yu
Key Lab of Aromatic Plant Resources Exploitation and Utilization in Sichuan Higher Education, Yibin College Yibin, Sichuan, China.
College of Veterinary Medicine Sichuan Agricultural University Ya-An 625014, China.
Int J Clin Exp Pathol. 2014 Sep 15;7(10):6524-33. eCollection 2014.
To investigate the effect of γ-terpineol on cell proliferation and apoptosis of human hepatoma BEL-7402 cells to elucidate its molecular mechanism. Here, BEL-7402 cells were treated with various concentrations (40, 80, 160, 320 and 640 μg/ml) of γ-terpineol for 48 h, cell proliferation was determined by 3-(4,5-dimethyl-thiazolyl-2)-2,5-diphenyl tetrazolium bromides (MTT) assay. Cell colony inhibition was determined by soft agar assay. Apoptosis and possible molecular mechanisms were evaluated by morphological observation, flow cytometry analysis, and DNA fragmentation assay. The γ-terpineol significantly suppressed BEL-7402 cell proliferation in a dose-dependent manner. Characteristic morphological and biochemical changes associated with apoptosis such as cells shrinkage, deformation and vacuolization of mitochondria, nuclear chromatin condensation and fragmentation, formation of apoptotic bodies were observed after BEL-7402 cells treated with γ-terpineol for 24 h and 48 h. Cell cycle were displayed by flow cytometry analysis, the γ-terpineol treatment resulted in accumulation of cells at G₁ or S phase and a blockade of cell proliferation compared to control group. Treating BEL-7402 cells with 320 μg/ml of γ-terpineol for 36 h and 48 h, a typical apoptotic "DNA ladder" was observed using DNA fragmentation assay. The present study demonstrated that possible anti-cancer mechanism of γ-terpineol on human hematomas cells is through inducing cell apoptosis to suppress tumor cell growth.
为研究γ-萜品醇对人肝癌BEL-7402细胞增殖和凋亡的影响,并阐明其分子机制。在此,用不同浓度(40、80、160、320和640μg/ml)的γ-萜品醇处理BEL-7402细胞48小时,通过3-(4,5-二甲基-噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法测定细胞增殖。通过软琼脂试验测定细胞集落抑制。通过形态学观察、流式细胞术分析和DNA片段化试验评估凋亡及可能的分子机制。γ-萜品醇以剂量依赖性方式显著抑制BEL-7402细胞增殖。在用γ-萜品醇处理BEL-7402细胞24小时和48小时后,观察到与凋亡相关的特征性形态和生化变化,如细胞收缩、线粒体变形和空泡化、核染色质浓缩和断裂、凋亡小体形成。通过流式细胞术分析显示细胞周期,与对照组相比,γ-萜品醇处理导致细胞在G₁期或S期积累并阻断细胞增殖。用320μg/ml的γ-萜品醇处理BEL-7402细胞36小时和48小时,使用DNA片段化试验观察到典型的凋亡“DNA梯带”。本研究表明,γ-萜品醇对人肝癌细胞可能的抗癌机制是通过诱导细胞凋亡来抑制肿瘤细胞生长。