Amantadine inhibits cellular proliferation and induces the apoptosis of hepatocellular cancer cells in vitro
Amantadine inhibits cellular proliferation and induces the apoptosis of hepatocellular cancer cells in vitro
复制标题
金刚烷胺在体外抑制细胞增殖并诱导肝细胞癌细胞凋亡。
DOI：
10.3892/ijmm.2015.2289
复制
发表时间：
2015-09-01
影响因子：
5.4
通讯作者：
Zhou, Jun
中科院分区：
文献类型：
--
作者：
Lan, Zengmei;Chong, Zhaoyang;Zhou, Jun
肝细胞癌（HCC）是世界范围内最具侵袭性的恶性肿瘤之一，近年来其与病毒感染相关的发病率呈上升趋势。金刚烷胺是一种三环对称胺，可以有效地防止丙型肝炎病毒。然而，其抗肿瘤特性仍不清楚。本研究探讨了金刚烷胺对肿瘤细胞活力、细胞周期调控和细胞凋亡的影响。MTT法检测肝癌细胞株HepG 2和SMMC-7721的生长情况。流式细胞仪检测细胞周期调控和凋亡。还进行了逆转录定量聚合酶链反应和蛋白质印迹分析，以检测细胞周期和凋亡相关基因和蛋白的表达，包括细胞周期蛋白E、细胞周期蛋白D1、细胞周期蛋白依赖性激酶2（CDK 2）、B细胞淋巴瘤2（Bcl-2）和Bax。我们的结果表明，金刚烷胺以剂量和时间依赖的方式显著抑制HepG 2和SMMC-7721细胞的增殖，并将细胞周期阻滞在G 0/G1期。金刚烷胺可降低细胞周期相关基因和蛋白（细胞周期蛋白D1、细胞周期蛋白E和CDK 2）的水平，并显著诱导细胞凋亡。金刚烷胺处理还降低了Bcl-2，增加了Bax蛋白和mRNA水平。此外，金刚烷胺处理后，两种HCC细胞系中的Bcl-2/Bax比值较低。总的来说，这些结果强调金刚烷胺在抑制肝癌细胞增殖和诱导细胞凋亡中的作用，提倡将其作为一种新的肿瘤抑制治疗候选药物。
Hepatocellular carcinoma (HCC) is one of the most aggressive malignancies worldwide, and its incidence associated with viral infection has increased in recent years. Amantadine is a tricyclic symmetric amine that can effectively protect against the hepatitis C virus. However, its antitumor properties remain unclear. In the present study, the effects of amantadine on tumor cell viability, cell cycle regulation and apoptosis were investigated. The growth of HepG2 and SMMC‑7721 cells (HCC cell lines) was detected by an MTT assay. Flow cytometry was used to investigate cell cycle regulation and apoptosis. Reverse transcription‑quantitative polymerase chain reaction and western blot analysis were also performed to examine the expression of cell cycle‑ and apoptosis‑related genes and proteins, including cyclin E, cyclin D1, cyclin‑dependent kinase 2 (CDK2), B‑cell lymphoma 2 (Bcl‑2) and Bax. Our results demonstrated that amantadine markedly inhibited the proliferation of HepG2 and SMMC‑7721 cells in a dose‑ and time‑dependent manner and arrested the cell cycle at the G0/G1 phase. The levels of the cell cycle‑related genes and proteins (cyclin D1, cyclin E and CDK2) were reduced by amantadine, and apoptosis was significantly induced. Amantadine treatment also reduced Bcl‑2 and increased the Bax protein and mRNA levels. Additionally, Bcl‑2/Bax ratios were lower in the two HCC cell lines following amantadine treatment. Collectively, these results emphasize the role of amantadine in suppressing proliferation and inducing apoptosis in HCC cells, advocating its use as a novel tumor-suppressive therapeutic candidate.