Liu Jie, Ren Hao, Liu Bin, Zhang Qingyu, Li Mingyi, Zhu Runzhi
Laboratory of Hepatobiliary Surgery, Affiliated Hospital of Guangdong Medical University, Zhanjiang Key Laboratory of Hepatobiliary Diseases, Zhanjiang, Guangdong 524001, P.R. China.
Oncol Lett. 2016 Dec;12(6):4385-4392. doi: 10.3892/ol.2016.5301. Epub 2016 Oct 19.
Hepatocellular carcinoma (HCC), which is a type of malignant tumor, is the fifth most common cancer in men and ninth in women worldwide. The aim of the present study was to investigate the antitumor effect of diosmetin (DIOS) in hepatocellular carcinoma HepG2 cells. The proliferation, apoptosis and autophagy rates of HepG2 cells were measured following treatment with DIOS. The effects of DIOS treatment on HepG2 cell proliferation and apoptosis rates were analyzed using MTT assays and Annexin V staining, respectively. The effect of DIOS treatment on autophagy levels was assessed using transmission electron microscopy, green fluorescent protein (GFP)-microtubule-associated protein 1 light chain (LC3) transfection and LysoTracker Red staining. Furthermore, bafilomycin A1 (BA1), an autophagy inhibitor, was used to assess the association between DIOS and cell autophagy, proliferation and apoptosis. In addition, the expression of autophagy-related proteins [mammalian target of rapamycin (mTOR), phosphatidylinositol 3-kinase, P70S6K, phosphoinositide-dependent kinase-1, extracellular signal-regulated kinase, 5'-AMP-activated protein kinase and Akt] and apoptosis-related proteins [B-cell lymphoma (Bcl)-2-associated X protein, Bak, p53, Bcl-2 and caspase-3] were analyzed by western blotting. The results revealed that DIOS significantly inhibited proliferation (P<0.01) and induced apoptosis (P<0.001) in HepG2 cells. It was also demonstrated that DIOS triggered autophagy by regulating the mTOR pathway in HepG2 cells. Notably, following treatment of HepG2 cells with the autophagy inhibitor, BA1, the expression of apoptosis-related proteins, including Bax, Bak and p53, were significantly decreased (P<0.05), and cell viability was recovered to a certain extent. In conclusion, DIOS inhibits cell proliferation and induces apoptosis in HepG2 cells via regulation of the mTOR pathway. Thus, the results of the current study indicate that DIOS may present a potential therapeutic agent for HCC treatment.
肝细胞癌(HCC)是一种恶性肿瘤,是全球男性中第五大常见癌症,女性中第九大常见癌症。本研究的目的是探讨香叶木素(DIOS)对肝癌HepG2细胞的抗肿瘤作用。用DIOS处理后,测定HepG2细胞的增殖、凋亡和自噬率。分别采用MTT法和膜联蛋白V染色分析DIOS处理对HepG2细胞增殖和凋亡率的影响。采用透射电子显微镜、绿色荧光蛋白(GFP)-微管相关蛋白1轻链(LC3)转染和溶酶体追踪染料Red染色评估DIOS处理对自噬水平的影响。此外,自噬抑制剂巴弗洛霉素A1(BA1)用于评估DIOS与细胞自噬、增殖和凋亡之间的关系。另外,通过蛋白质免疫印迹法分析自噬相关蛋白[雷帕霉素靶蛋白(mTOR)、磷脂酰肌醇3激酶、P70S6K、磷酸肌醇依赖性激酶-1、细胞外信号调节激酶、5'-AMP激活蛋白激酶和Akt]和凋亡相关蛋白[B细胞淋巴瘤(Bcl)-2相关X蛋白、Bak、p53、Bcl-2和半胱天冬酶-3]的表达。结果显示,DIOS显著抑制HepG2细胞的增殖(P<0.01)并诱导其凋亡(P<0.001)。还证明DIOS通过调节HepG2细胞中的mTOR途径触发自噬。值得注意的是,用自噬抑制剂BA1处理HepG2细胞后,包括Bax、Bak和p53在内的凋亡相关蛋白的表达显著降低(P<0.05),细胞活力在一定程度上得以恢复。总之,DIOS通过调节mTOR途径抑制HepG2细胞的增殖并诱导其凋亡。因此,本研究结果表明DIOS可能是一种潜在的肝癌治疗药物。