S腺苷甲硫氨酸对肝癌细胞急性缺血/缺氧过程中的生物学作用  

Effect of S-adenosylmethionine on hepatic cancer cell during acute ischemia/hypoxia

作　　者：佟辉[1] 杨卫平[1] 林大伟[1] 施敏敏[1] 沈柏用[1] 彭承宏[1] 李宏为[1] 邱伟华[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院外科 上海消化外科研究所,上海200025

出　　处：《外科理论与实践》2013年第5期435-439,共5页Journal of Surgery Concepts & Practice

基　　金：国家自然科学基金(81172326);上海市自然科学基金(10ZR1419400);上海慈善癌症研究基金

摘　　要：目的:研究S腺苷甲硫氨酸(SAMe)在急性缺血/缺氧过程中对肝癌细胞HepG2的生物学作用,并通过检测自噬的变化探讨其可能机制。方法:real time-PCR检测自噬特异性基因(Beclin 1)表达的变化;吖啶橙染色后,采用荧光显微镜定性观察自噬;CCK-8法检测HepG2细胞的成活率;用AnnexinⅤ/PI流式细胞仪检测细胞凋亡的变化。结果:SAMe可诱导HepG2细胞的自噬,在单纯缺血和缺血/缺氧处理因素下,HepG2自噬在荧光强度和Beclin 1基因水平分别比空白对照组增强约3.2倍和3.5倍。细胞成活率分别比空白对照组增加了约20%和30%。SAMe对HepG2细胞具有显著的抑制作用,这种抑制作用在缺血/缺氧环境中更加明显,SAMe单独处理组和SAMe预处理结合缺血/缺氧组,其细胞成活率与空白对照组相比分别下降了约30%和70%。随着细胞成活率下降,细胞凋亡比例相应增加,与空白对照组相比,细胞经SAMe处理后凋亡比例增加约18%;细胞经SAMe预处理后再予以缺血/缺氧,凋亡比例增加约30%。结论:在SAMe抑制HepG2细胞生长过程中,自噬起着重要作用。Objective To investigate the role of S-adenosylmethionine （SAMe） during acute ischemidhypoxia of hepatic cancer cell and its mechanism by analysis of autophagy. Methods The mRNA expression of Beclin 1 was measured by real time PCR. Quantitative analysis of autophagy was performed by fluorescent microscope on acridine orange staining. The cellular viability was detected by the CCK-8 method. The apoptosis was detected by Annexin V/PI flow cytometry. Results SAMe induced autophagy in hepatoma cell line HepG2. The cellular viability under ischemia and ischemia/hypoxia respectively increased about 20% and 30%. The level of autophagy in fluorescence intensity and mRNA of Beclin 1 expression increased about 3.2 and 3.5 times. SAMe significantly inhibited the proliferation of HepG2, especially in the environment of ischemia/hypoxia. The cellular viability under SAMe treatment only and SAMe-treatment followed by ischemia/hypoxia decreased about 30% and 70%, respectively. The cellular apoptosis increased about 18% and 30% compared to the blank control. Conclusions SAMe inhibits the proliferation of HepG2, in which autophagy may play an important role.

关 键 词：肝癌 自噬 S腺苷甲硫氨酸 

分 类 号：R735.7[医药卫生—肿瘤]