Wang Zhen, Ying You-Min, Li Kai-Qiang, Zhang Yu, Chen Bing-Yu, Zeng Jing-Jing, He Xu-Jun, Jiang Meng-Meng, Chen Bo-Xu, Wang Ying, Xu Xiao-Dong, Hao Ke, Zhu Meng-Hua, Zhang Wei
Research Center of Blood Transfusion Medicine, Education Ministry Key Laboratory of Laboratory Medicine, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou, Zhejiang 310014, P.R. China.
School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China.
Exp Ther Med. 2017 Sep;14(3):2477-2484. doi: 10.3892/etm.2017.4833. Epub 2017 Jul 25.
An extract from a traditional Chinese herb, (trade name, Xiao-Ai-Ping) has been approved for use on the Chinese market as a cancer chemotherapeutic agent for decades. Previous studies have demonstrated the cytostatic and pro-apoptotic effects of extract (MTE) in multiple cancer cells. However, the contributions of MTE to the proliferation and apoptosis of hepatoma carcinoma cells and the underlying mechanisms remain unclear. In the present study, Bel-7402 cells were incubated with increasing concentrations of MTE ranging from 0-320 µl/ml to explore the effects and potential mechanisms of MTE on the proliferation and apoptosis of Bel-7402 cells. 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium, inner salt and propidium iodide (PI)-stained flow cytometry assays demonstrated that MTE significantly suppressed the proliferation of Bel-7402 cells in a dose-dependent manner by arresting the cell cycle at S phase (P<0.05). Annexin V-fluorescein isothiocyanate PI-stained flow cytometry confirmed the significantly pro-apoptotic effect of MTE at both 160 and 240 µl/ml (P<0.001). Reverse transcription-quantitative polymerase chain reaction and western blot analysis demonstrated that MTE (both 160 and 240 µl/ml) induced a significant downregulation of B-cell lymphoma (Bcl)-2 (P<0.01), upregulation of Bcl-2-associated X protein (P<0.01) and activation of caspase-3 (P<0.05). Furthermore, a significant downregulation of murine double minute-2 (MDM2) (P<0.001) and activation of p53 (P<0.001) in Bel-7402 cells following treatment with 160 or 240 µl/ml MTE was observed, accompanied by the inhibition of the nuclear factor (NF)-κB pathway (P<0.001). These results suggested that MTE inhibited growth and exhibited pro-apoptotic effects in Bel-7402 cells, which was mediated by downregulation of the MDM2-induced p53-dependent mitochondrial apoptosis pathway and blocking the NF-κB pathway. Overall, these data serve as preliminary identification of the significant roles of MTE in hepatic carcinoma cells, and suggest that MTE may be a promising candidate for hepatocellular carcinoma therapy.
一种源自传统中草药的提取物（商品名，消癌平）数十年来已在中国市场获批用作癌症化疗药物。先前的研究已证明该提取物（MTE）在多种癌细胞中具有抑制细胞生长和促凋亡作用。然而，MTE对肝癌细胞增殖和凋亡的作用及其潜在机制仍不清楚。在本研究中，将Bel - 7402细胞与浓度范围为0 - 320 μl/ml递增的MTE孵育，以探究MTE对Bel - 7402细胞增殖和凋亡的影响及潜在机制。3 -（4,5 - 二甲基噻唑 - 2 - 基）- 5（3 - 羧甲氧基苯基）- 2 -（4 - 磺基苯基）- 2H - 四唑鎓内盐和碘化丙啶（PI）染色的流式细胞术检测表明，MTE通过使细胞周期停滞在S期以剂量依赖性方式显著抑制Bel - 7402细胞的增殖（P<0.05）。膜联蛋白V - 异硫氰酸荧光素PI染色的流式细胞术证实，160和240 μl/ml的MTE均具有显著的促凋亡作用（P<0.001）。逆转录定量聚合酶链反应和蛋白质印迹分析表明，MTE（160和240 μl/ml）均导致B细胞淋巴瘤（Bcl）- 2显著下调（P<0.01），Bcl - 2相关X蛋白上调（P<0.01）以及半胱天冬酶 - 3激活（P<0.05）。此外，在用160或240 μl/ml MTE处理后的Bel - 7402细胞中，观察到鼠双微体2（MDM2）显著下调（P<0.001）和p53激活（P<0.001），同时伴有核因子（NF）-κB通路的抑制（P<0.001）。这些结果表明，MTE在Bel - 7402细胞中抑制生长并表现出促凋亡作用这一过程是由MDM2诱导的p53依赖性线粒体凋亡通路的下调以及NF - κB通路的阻断介导的。总体而言，这些数据初步确定了MTE在肝癌细胞中的重要作用，并表明MTE可能是肝细胞癌治疗的一个有前景的候选药物。
