Plumbagin induces autophagy and apoptosis of SMMC-7721 cells in vitro and in vivo
Plumbagin induces autophagy and apoptosis of SMMC-7721 cells in vitro and in vivo
复制标题
白花丹素在体外和体内诱导 SMMC-7721 细胞自噬和凋亡
DOI：
10.1002/jcb.28262
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发表时间：
2019-06-01
影响因子：
4
通讯作者：
Wei, Yanfei
中科院分区：
文献类型：
--
作者：
Lin, Yuning;Chen, Yongxin;Wei, Yanfei
白花丹素（Plumbagin，PL）是一种具有生物活性的萘醌类化合物，具有潜在的抗癌活性.然而，潜在的抗癌机制尚未完全了解。在这项研究中，人肝癌（HCC）SMMC-7721细胞系进行了研究，在体外模型。PL对细胞增殖有明显的抑制作用，且呈时间和剂量依赖性。采用电镜、吖啶橙子染色和免疫荧光法观察PL处理SMMC-7721细胞后自噬体的形成和LC 3蛋白的表达。实时荧光定量PCR和Western blot结果显示PL处理抑制了SMMC-7721细胞中与肿瘤凋亡和自噬相关的凋亡和自噬因子（LC 3、Beclin 1、Atg 7和Atg 5）的表达。在体外裸鼠肿瘤模型的研究中，PL能抑制肿瘤生长。采用苏木精-伊红染色、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记染色和Western blot检测移植瘤细胞凋亡和自噬。此外，在肝癌细胞的体内研究中，我们发现用自噬抑制剂3-甲基腺嘌呤预处理可以阻断PL诱导的凋亡的形成。相反，凋亡抑制剂Z-VAD的管理并不影响PL诱导的自噬。综上所述，我们的研究结果强烈表明PL是一种有前途的药物，具有显着的抗肝癌活性。
Plumbagin (PL), an active naphthoquinone compound, has been demonstrated to be a potential anticancer agent. However, the underlying anticancer mechanism is not fully understood. In this study, the human hepatocellular carcinoma (HCC) SMMC-7721 cell line was studied in an in vitro model. The cell proliferation was inhibited by PL in a dose- and time-dependent manner. Electron microscopy, acridine orange staining, and immunofluorescence were used to evaluate autophagosome formation and LC3 protein expression in PL-treated SMMC-7721 cells. Real-time polymerase chain reaction and Western blot showed that PL treatment suppressed the expression of apoptosis and autophagy factors (LC3, Beclin1, Atg7, and Atg5), which are associated with tumor apoptosis and autophagy in SMMC-7721 cells. In the study of in vitro tumor nude mouse models, PL can inhibit tumor growth. Cell apoptosis and autophagy of the transplanted tumors were evaluated by hematoxylin and eosin staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling staining, and Western blot. In addition, in the in vivo studies of HCC cells, we found that pretreatment with the autophagy inhibitor 3-methyladenine blocked the formation of apoptosis induced by PL. In contrast, administration of the apoptosis inhibitor Z-VAD did not affect PL-induced autophagy. Taken together, our findings strongly suggest that PL is a promising drug with significant antitumor activity in HCC.