Zhang Jixiang, Wu Dandan, Song Jia, Wang Jing, Yi Jiasheng, Dong Weiguo
Department of Gastroenterology, Renmin Hospital of Wuhan University, Wuhan, 430060, Hubei Province, China.
Dig Dis Sci. 2015 Oct;60(10):2985-95. doi: 10.1007/s10620-015-3696-7. Epub 2015 May 14.
Hesperetin, has been shown to exert biological activities on various types of human cancers. However, few related studies on gastric cancer are available.
In this study, we sought to investigate the effect of hesperetin on gastric cancer and clarify its specific mechanism.
Cell Counting Kit-8, 2',7'-dichlorofluorescin diacetate, JC-1, Hoechst 33258 staining, and western bolt were used to detect cell viability, levels of intracellular reactive oxygen species (ROS), changes in mitochondrial membrane potential (△ψ m), cell apoptosis, and expressions of mitochondrial pathway proteins, respectively. Meanwhile, xenograft tumor models in nude mice were made to evaluate the effect of hesperetin on gastric cancer in vivo.
Compared with the control group, the proliferation of gastric cancer cells in hesperetin groups was significantly inhibited (P < 0.05), and dose- and time-dependent effects were observed. Pretreatment with H2O2 (1 mM) or N-acetyl-L-cysteine (5 mM) enhanced or attenuated the hesperetin-induced inhibition of cell viability (P < 0.05). Percentages of apoptotic cells, levels of intracellular ROS, and △ψ m varied with the dose and treatment time of hesperetin (P < 0.05), and hesperetin caused an increase in the levels of AIF, Apaf-1, Cyt C, caspase-3, caspase-9, and Bax and a decrease in Bcl-2 levels (P < 0.05). Meanwhile, hesperetin significantly inhibited the growth of xenograft tumors (P < 0.05).
Our study suggests that hesperetin could inhibit the proliferation and induce the apoptosis of gastric cancer cells via activating the mitochondrial pathway by increasing the ROS.
橙皮素已被证明对多种类型的人类癌症具有生物学活性。然而，关于胃癌的相关研究较少。
在本研究中，我们试图研究橙皮素对胃癌的影响并阐明其具体机制。
分别使用细胞计数试剂盒-8、2',7'-二氯荧光素二乙酸酯、JC-1、Hoechst 33258染色和蛋白质免疫印迹法检测细胞活力、细胞内活性氧（ROS）水平、线粒体膜电位（△ψm）变化、细胞凋亡以及线粒体途径蛋白的表达。同时，建立裸鼠异种移植瘤模型以评估橙皮素在体内对胃癌的影响。
与对照组相比，橙皮素组胃癌细胞的增殖受到显著抑制（P < 0.05），并观察到剂量和时间依赖性效应。用H2O2（1 mM）或N-乙酰半胱氨酸（5 mM）预处理增强或减弱了橙皮素诱导细胞活力的抑制作用（P < 0.05）。凋亡细胞百分比、细胞内ROS水平和△ψm随橙皮素的剂量和处理时间而变化（P < 0.05），橙皮素导致AIF、Apaf-1、Cyt C、caspase-3、caspase-9和Bax水平升高，Bcl-2水平降低（P < 0.05）。同时，橙皮素显著抑制异种移植瘤的生长（P < 0.05）。
我们的研究表明，橙皮素可通过增加ROS激活线粒体途径来抑制胃癌细胞的增殖并诱导其凋亡。
