Lee Seung-On, Joo Sang Hoon, Kwak Ah-Won, Lee Mee-Hyun, Seo Ji-Hye, Cho Seung-Sik, Yoon Goo, Chae Jung-Il, Shim Jung-Hyun
Department of Biomedicine, Health & Life Convergence Sciences, BK21 Four, Biomedical and Healthcare Research Institute, Mokpo National University, Muan 58554, Republic of Korea.
College of Pharmacy, Daegu Catholic University, Gyeongsan 38430, Republic of Korea.
Biomol Ther (Seoul). 2021 Nov 1;29(6):658-666. doi: 10.4062/biomolther.2021.143.
Podophyllotoxin (PT), a lignan compound from the roots and rhizomes of , has diverse pharmacological activities including anticancer effect in several types of cancer. The molecular mechanism of the anticancer effects of PT on colorectal cancer cells has not been reported yet. In this study, we sought to evaluate the anticancer effect of PT on human colorectal cancer HCT116 cells and identify the detailed molecular mechanism. PT inhibited the growth of cells and colony formation in a concentration-dependent manner and induced apoptosis as determined by the annexin V/7-aminoactinomycin D double staining assay. PT-induced apoptosis was accompanied by cell cycle arrest in the G2/M phase and an increase in the generation of reactive oxygen species (ROS). The effects of PT on the induction of ROS and apoptosis were prevented by pretreatment with N-acetyl-L-cysteine (NAC), indicating that an increase in ROS generation mediates the apoptosis of HCT116 cells induced by PT. Furthermore, Western blot analysis showed that PT upregulated the level of phospho (p)-p38 mitogen-activated protein kinase (MAPK). The treatment of SB203580, a p38 inhibitor, strongly prevented the apoptosis induced by PT, suggesting that PT-induced apoptosis involved the p38 MAPK signaling pathway. In addition, PT induced the loss of mitochondrial membrane potential and multi-caspase activation. The results suggested that PT induced cell cycle arrest in the G2/M phase and apoptosis through the p38 MAPK signaling pathway by upregulating ROS in HCT116 cells.
鬼臼毒素(PT)是一种从[植物名称]的根和根茎中提取的木脂素化合物,具有多种药理活性,包括对多种癌症的抗癌作用。PT对结肠癌细胞抗癌作用的分子机制尚未见报道。在本研究中,我们试图评估PT对人结肠癌细胞HCT116的抗癌作用,并确定其详细的分子机制。PT以浓度依赖的方式抑制细胞生长和集落形成,并通过膜联蛋白V/7-氨基放线菌素D双染法检测诱导细胞凋亡。PT诱导的细胞凋亡伴随着细胞周期阻滞在G2/M期和活性氧(ROS)生成增加。用N-乙酰-L-半胱氨酸(NAC)预处理可阻止PT对ROS诱导和细胞凋亡的影响,表明ROS生成增加介导了PT诱导的HCT116细胞凋亡。此外,蛋白质印迹分析表明PT上调了磷酸化(p)-p38丝裂原活化蛋白激酶(MAPK)的水平。p38抑制剂SB203580的处理强烈阻止了PT诱导的细胞凋亡,提示PT诱导的细胞凋亡涉及p38 MAPK信号通路。此外,PT诱导线粒体膜电位丧失和多胱天蛋白酶激活。结果表明,PT通过上调HCT116细胞中的ROS,通过p38 MAPK信号通路诱导细胞周期阻滞在G2/M期和细胞凋亡。