Bagheri E, Hajiaghaalipour F, Nyamathulla S, Salehen N A
Department of Pharmacy, Faculty of Medicine, University of Malaya 50603 Kuala Lumpur Malaysia
Institute of Biological Science, Faculty of Science, University of Malaya 50603 Kuala Lumpur Malaysia.
RSC Adv. 2018 Jan 2;8(2):681-689. doi: 10.1039/c7ra09618f.
(L.) Merr. is a well-known plant in Chinese System of Medicine. Its fruits and seeds have been reported to possess curative properties against various ailments. The chemical constituents and biological activity of this plant have been an interesting area in plant and chemistry medicine. The aim of this study is to evaluate the antiproliferative effects of the extract against a colon cancer cell line and identification of the chemical components derived from the extract. An ethanolic extract from fruits was prepared by cold maceration method, subjected to LC-MS profiling to elucidate the composition abbreviated as BJEE. The extract was screened for the cytotoxicity effects on HCT-116 colon cancer cells MTT and LDH methods. Additionally, AO/PI staining verified apoptosis features in HCT-116 cells through microscopic analysis. ROS, caspase activity, and gene expression has been performed to identify its possible mechanism of actions which contribute to apoptosis. Output data from this study showed BJEE inhibited the cell proliferation of HCT-116 colon cancer cells at IC value of 8.9 ± 1.32 (μg mL) and significantly increased the levels of caspase-8, 9, and 3/7 in treated cells in comparison to untreated. The changes in expression of caspase genes and some apoptosis genes like Bax and Bcl-2 were confirmed using RT-PCR. Phytochemical analysis by LC-MS identified six major active compounds (bruceine D, isobrucein A, quassimarin, C16 sphinganine, phytosphingosine, and enigmol) in BJEE that may play a key role in cell apoptosis. The current study showed BJEE could be a promising agent for colorectal cancer therapy by significant increase in caspase activity level, and up-regulation of the specific apoptotic genes.
（L.）Merr.是中国医学体系中一种著名的植物。据报道，其果实和种子具有治疗多种疾病的特性。该植物的化学成分和生物活性一直是植物医学和化学医学领域一个有趣的研究方向。本研究的目的是评估该提取物对结肠癌细胞系的抗增殖作用，并鉴定提取物中的化学成分。通过冷浸法制备了（植物名称缺失）果实的乙醇提取物，进行液相色谱 - 质谱分析以阐明其成分，简称为BJEE。采用MTT法和LDH法筛选该提取物对HCT - 116结肠癌细胞的细胞毒性作用。此外，通过AO/PI染色，经显微镜分析验证HCT - 116细胞中的凋亡特征。还进行了活性氧（ROS）、半胱天冬酶活性和基因表达分析，以确定其导致细胞凋亡的可能作用机制。本研究的输出数据表明，BJEE在IC值为8.9±1.32（μg/mL）时抑制HCT - 116结肠癌细胞的增殖，与未处理细胞相比，显著提高了处理细胞中半胱天冬酶 - 8、9和3/7的水平。使用逆转录 - 聚合酶链反应（RT - PCR）确认了半胱天冬酶基因以及一些凋亡相关基因如Bax和Bcl - 2表达的变化。通过液相色谱 - 质谱进行的植物化学分析在BJEE中鉴定出六种主要活性化合物（鸦胆子丁、异鸦胆子素A、苦木素、C16鞘氨醇、植物鞘氨醇和enigmol），它们可能在细胞凋亡中起关键作用。当前研究表明，BJEE通过显著提高半胱天冬酶活性水平和上调特定凋亡基因，可能成为结直肠癌治疗的一种有前景的药物。
