Sinomenine hydrochloride inhibits human hepatocellular carcinoma cell growth in vitro and in vivo: Involvement of cell cycle arrest and apoptosis induction
Sinomenine hydrochloride inhibits human hepatocellular carcinoma cell growth in vitro and in vivo: Involvement of cell cycle arrest and apoptosis induction
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DOI：
10.3892/ijo.2012.1704
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发表时间：
2013-01-01
影响因子：
5.2
通讯作者：
He, Shui-Xiang
中科院分区：
文献类型：
--
作者：
Lu, Xin-Lan;Zeng, Jin;He, Shui-Xiang
肝细胞癌（HCC）是世界范围内最常见的恶性肿瘤之一。然而，迄今为止，针对HCC的治疗尚未完全有效。盐酸青藤碱（Sinomenine hydrochloride，SH）是一种临床应用的抗关节炎药物，在体外对多种肿瘤细胞具有抗肿瘤活性。SH是否抑制HCC仍不清楚。为此，在本研究中，MTT法用于测定细胞生长。采用流式细胞术、Hoechst染色、DNA片段化、Western blot分析、免疫组化和TUNEL染色等方法研究其作用机制。使用小鼠异种移植模型测定SH的体内活性。SH在体外可抑制多种类型的人肝癌细胞的生长。我们发现，SH促进细胞周期停滞在G1期和亚G1期的形成，与增加p21/WAF 1/Cip 1的表达。此外，SH诱导caspase依赖性凋亡，包括线粒体膜电位的破坏，细胞色素c和Omi/HtrA 2从线粒体释放到细胞质中的增加，Bcl-2的下调和Bax的上调，caspase级联（caspase-8，-10，-9和-3）和PARP的激活，以及Survivin表达的降低。SH抑制人肝癌异种移植瘤生长是由于细胞增殖减少和诱导细胞凋亡，涉及caspase-3的激活，p21的上调和Survivin的下调。这些结果表明，SH在体外和体内表现出抗癌功效，涉及细胞周期和半胱天冬酶依赖性凋亡，并可能作为一个潜在的候选药物抗肝癌。
Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide. However, therapies against HCC to date have not been completely effective. Sinomenine hydrochloride (SH), an anti-arthritis drug applied in clinical practice, has been reported to have in vitro anti-neoplastic activity in various cancer cells. Whether SH inhibits HCC remains unknown. For this purpose, in this study, MTT assay was used to determine cell growth. Flow cytometry, Hoechst staining, DNA fragmentation, western blot analysis, immunohistochemisty and TUNEL staining were performed to investigate the mechanisms involved. The in vivo activity of SH was determined using a mouse xenograft model. SH inhibited the growth of various types of human HCC cells in vitro. We found that SH promoted cell cycle arrest in the G1 phase and sub-G1 formation, associated with the increased p21/WAF1/Cip1 expression. Additionally, SH induced caspase-dependent apoptosis, which involved the disruption of mitochondrial membrane potential, the increased release of cytochrome c and Omi/HtrA2 from the mitochondria into the cytoplasm, the downregulation of Bcl-2 and the upregulation of Bax, the activation of a caspase cascade (caspase-8, -10, -9 and -3) and PARP, as well as the decreased expression of survivin. The SH-suppressed growth of human HCC xenografts in vivo occurred due to the decrease in proliferation and the induction of apoptosis, implicating the activation of caspase-3, the upregulation of p21 and the downregulation of survivin. These findings suggest that SH exhibits anticancer efficacy in vitro and in vivo involving cell cycle and caspase-dependent apoptosis and may serve as a potential drug candidate against HCC.