Icaritin enhances sorafenib-induced apoptosis through a mitochondria-dependent pathway  

淫羊藿素通过线粒体通路增强索拉非尼的诱导凋亡作用

作　　者：Axi Shi Tiantian Shen Wenbin Xia Lili Xi Lijun Wang Yuhui Wei 石阿茜;沈阗阗;夏文彬;席莉莉;王利军;魏玉辉(兰州大学第一医院药剂科,甘肃兰州730000;兰州大学第一临床医学院,甘肃兰州730000;兰州大学药学院,甘肃兰州730000;兰州大学第一医院国家药物临床试验机构办公室,甘肃兰州730000)

机构地区：[1]Department of Pharmacy,the First Hospital of Lanzhou University,Lanzhou 730000,China [2]The First Clinical Medical College,Lanzhou University,Lanzhou 730000,China [3]School of Pharmacy,Lanzhou University,Lanzhou 730000,China [4]Office of the National Institute for Clinical Trials of Drugs,First Hospital of Lanzhou University,Lanzhou 730000,China

出　　处：《Journal of Chinese Pharmaceutical Sciences》2022年第12期928-937,共10页中国药学（英文版）

摘　　要：Sorafenib remains the standard systemic treatment for advanced human hepatocellular carcinoma(HCC).However,the low response rate,high recurrence,and high progression limit the therapeutic efficacy.Therefore,a combination therapy strategy was advanced to strengthen the antitumor effects of sorafenib.In the present study,we aimed to evaluate whether icaritin could enhance the inhibitory effects of sorafenib on HCC cells and clarify the underlying mechanism.The cell viability was evaluated via MTT assay,and the synergistic inhibitory effects of sorafenib and icaritin were verified by calculating the combination index(CI).Their combined effects on cell proliferation or apoptosis were investigated using colony formation assay and flow cytometry.Mitochondrial membrane potential(MMP)was detected by flow cytometric assay.The protein expressions associated with the apoptotic pathway were determined by Western blotting analysis.The data demonstrated that sorafenib and icaritin exerted synergistic inhibitory effects on cell viability(CI<1).Icaritin enhanced the inhibitory effect of sorafenib on colony formation and sorafenib-induced apoptosis of HCC cells.We discovered a reduced level of antiapoptotic Bcl-2 and an elevated level of proapoptotic protein Bax when the cells were exposed to the combination.The effect of cleaved and activated PARP was also enhanced.Cleaved caspase-9 and cleaved caspase-3 were increased markedly in the combination group.Furthermore,the combination of icaritin and sorafenib significantly increased the loss of MMP compared with the single treatment group and induced the release of cytochrome c from the mitochondria to the cytosol.These findings indicated that icaritin could enhance sorafenib-induced cytotoxicity and trigger sorafenib-induced apoptosis through a mitochondria-dependent pathway.索拉非尼是晚期人肝细胞癌(human hepatocellular carcinoma,HCC)的标准全身治疗药物,但在治疗中出现的低应答率、高复发率和高进展限制了其疗效。因此,以加强索拉非尼的抗癌作用为目的的联合其他药物的治疗策略被广泛研究。本研究的目的是考察淫羊藿素增强索拉非尼对人肝癌细胞的抑制作用,并阐明其潜在机制。采用MTT法检测细胞存活率,并通过计算联合用药指数(combination index,CI)确定索拉非尼和淫羊藿素是否对细胞生长具有协同抑制作用。细胞克隆形成试验和流式细胞术分别考察药物对细胞增殖和凋亡的影响,采用流式细胞术检测线粒体膜电位(mitochondrial membrane potential,MMP),通过western blot技术分析凋亡通路相关蛋白的表达。结果显示,索拉非尼联合淫羊藿素对肝癌细胞的生长具有协同抑制作用(CI<1)。淫羊藿素一方面可增强索拉非尼对肝细胞集落形成的抑制作用,另一方面可增加索拉非尼诱导的肝细胞凋亡。联合治疗后,抗凋亡蛋白Bcl-2表达降低,而促凋亡蛋白Bax表达升高。PARP、caspase-9和caspase-3的剪切型表达显著增加。此外,与单用索拉非尼相比,联合用药使线粒体膜电位进一步丢失,进而导致线粒体通透性发生变化,细胞色素c从线粒体释放到胞质中。结果表明,淫羊藿素可增加索拉非尼诱导的细胞毒作用,并通过线粒体依赖性途径增加索拉非尼诱导的细胞凋亡。

关 键 词：SORAFENIB ICARITIN Human hepatocellular carcinoma 

分 类 号：R966[医药卫生—药理学]