Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China.
Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, Central Laboratory, Peking University Cancer Hospital and Institute, Beijing 100142, P.R. China.
Mol Med Rep. 2018 Aug;18(2):2068-2078. doi: 10.3892/mmr.2018.9203. Epub 2018 Jun 21.
The management of patients with triple‑negative breast cancer is challenging due to the lack of effective therapeutic options, aggressive behavior and relatively poor prognosis. Xi Huang pills (XHP) are a well‑known traditional Chinese medicine that demonstrate anticancer activities. The aim of the present study was to investigate the antitumor effects of XHP on MDA‑MB‑231 cells in vitro and in vivo, and its potential underlying molecular mechanisms. In the present study, an MTT assay was used to evaluate the antiproliferative activity of XHP on MDA‑MB‑231 cells. In order to investigate the effects further, cell cycle distribution, apoptosis and mitochondrial membrane potential assays were performed, as well as western blot analyses. In addition, a tumor xenograft model was employed to investigate the effects of XHP in vivo. The results of the MTT assay demonstrated that the viability of MDA‑MB‑231 cells was markedly inhibited by XHP in a dose‑ and time‑dependent manner. The inhibitory effect of XHP on the viability of MDA‑MB‑231 cells was greater when compared with MCF‑10A cells. An increase in apoptosis and loss of mitochondrial membrane potential was observed following 4, 8 and 12 mg/ml XHP treatment of MDA‑MB‑231 cells. The protein expression levels of cleaved caspase‑3 were increased by 1.62‑, 2.13‑ and 2.19‑fold, respectively, when compared with the untreated controls, whereas no effects on the expression of B‑cell lymphoma 2 (Bcl‑2) or Bcl‑2‑associated X protein (Bax) were observed. The results of the cell cycle distribution assay analysis demonstrated that XHP treatment arrested cells at the G2/M phase. In addition, XHP treatment decreased the expression of cyclin A and increased the expression of p21Cip1. In vivo experiments revealed that XHP inhibited the growth of MDA‑MB‑231 xenograft tumors without body weight loss, and demonstrated similar effects on the protein expression levels of cleaved caspase 3, cyclin A and p21Cip1 as observed in vitro. In conclusion, the viability of MDA‑MB‑231 cells was inhibited by XHP in a dose‑dependent, time‑dependent and cell‑selective manner in vitro, and the potential underlying mechanisms may involve apoptosis and cell cycle arrest at the G2/M phase. XHP may induce apoptosis in MDA‑MB‑231 cells via the intrinsic pathway, which does not involve the Bcl‑2/Bax ratio. G2/M phase arrest may have been due to the integrated action of decreased cyclin A expression and increased p21Cip1 expression. In addition, XHP inhibited the growth of xenograft tumors in the absence of body weight loss in vivo.
由于缺乏有效的治疗选择、侵袭性行为和相对较差的预后，三阴性乳腺癌患者的管理具有挑战性。喜黄丸（XHP）是一种著名的中药，具有抗癌活性。本研究旨在探讨 XHP 在体外和体内对 MDA-MB-231 细胞的抗肿瘤作用及其潜在的分子机制。在本研究中，采用 MTT 法评估 XHP 对 MDA-MB-231 细胞的增殖抑制活性。为了进一步研究其作用机制，进行了细胞周期分布、细胞凋亡和线粒体膜电位测定以及 Western blot 分析。此外，还构建了肿瘤异种移植模型以研究 XHP 在体内的作用。MTT 检测结果表明，XHP 呈剂量和时间依赖性地显著抑制 MDA-MB-231 细胞的活力。XHP 对 MDA-MB-231 细胞活力的抑制作用大于 MCF-10A 细胞。XHP 处理 MDA-MB-231 细胞 4、8 和 12mg/ml 后，观察到凋亡增加和线粒体膜电位丧失。与未处理对照组相比，Cleaved caspase-3 的蛋白表达水平分别增加了 1.62、2.13 和 2.19 倍，而 B 细胞淋巴瘤 2（Bcl-2）或 Bcl-2 相关 X 蛋白（Bax）的表达无变化。细胞周期分布分析结果表明，XHP 处理将细胞阻滞在 G2/M 期。此外，XHP 处理降低了细胞周期蛋白 A 的表达，增加了 p21Cip1 的表达。体内实验结果表明，XHP 抑制 MDA-MB-231 异种移植肿瘤的生长，而无体重减轻，并且在体外观察到的 cleaved caspase 3、细胞周期蛋白 A 和 p21Cip1 的蛋白表达水平也有类似的作用。综上所述，XHP 在体外以剂量依赖性、时间依赖性和细胞选择性方式抑制 MDA-MB-231 细胞的活力，潜在机制可能涉及细胞凋亡和 G2/M 期细胞周期阻滞。XHP 可能通过内在途径诱导 MDA-MB-231 细胞凋亡，而不涉及 Bcl-2/Bax 比值。G2/M 期阻滞可能是由于细胞周期蛋白 A 表达降低和 p21Cip1 表达增加的综合作用。此外，XHP 在体内抑制异种移植肿瘤的生长而不引起体重减轻。
