Department of Oncology, Yantai Hospital of Traditional Chinese Medicine, Shandong, 264001, China.
Department of Clinical Laboratory, Yantai Hospital of Traditional Chinese Medicine, Shandong, 264001, China.
Cell Mol Biol (Noisy-le-grand). 2022 May 31;68(5):153-160. doi: 10.14715/cmb/2022.68.5.21.
The study aimed to explore the effects of atractylon on the proliferation and apoptosis of intestinal cancer cells through the phosphatidylinositol 3-hydroxy kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway. The intestinal cancer HT29 cell lines were cultured in vitro, and atractylon at different concentrations (15 and 30 mg/mL) was added. Then cell proliferative activity was detected via cell counting kit-8 (CCK8) assay, and the proportion of positive cells was determined using EdU staining. The content of interferon-γ (INF-γ), tumor necrosis factor-α (TNF-α) and matrix metalloproteinase-9 (MMP-9) was detected via enzyme-linked immunosorbent assay (ELISA), and the apoptosis of HT29 cells was detected through terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Moreover, reverse transcription-polymerase chain reaction (RT-PCR) was performed to determine the messenger ribonucleic acid (mRNA) levels of proliferation, apoptosis and PI3K/AKT/mTOR signaling pathway-related genes, and Western blotting was used to analyze the expression of the PI3K/AKT/mTOR signaling pathway. The cell growth status was poorer with a lower density in the 15 mg/mL atractylon group and basically normal morphological structure in the 30 mg/mL atractylon group. The number of cells significantly declined and the proliferative activity was also significantly weakened in the 30 mg/mL atractylon group. There were obviously more apoptotic cells in the 30 mg/mL atractylon group. Besides, INF-γ, TNF-α and MMP-9 were all evidently decreased in the 30 mg/mL atractylon group. Expressions of B-cell lymphoma-2 (Bcl-2), PI3K, AKT and mTOR obviously declined in the 30 mg/mL atractylon group, and they were raised in the NC group, while the expression of Caspase3 showed the opposite trends. Atractylon at an appropriate concentration can inhibit the proliferation and promote the apoptosis of intestinal cancer cells by suppressing the PI3K/AKT/mTOR signaling pathway, which can be used to treat colorectal cancer and other related diseases.
本研究旨在探讨苍术内酯通过磷脂酰肌醇 3-羟激酶（PI3K）/蛋白激酶 B（AKT）/哺乳动物雷帕霉素靶蛋白（mTOR）信号通路对肠癌细胞增殖和凋亡的影响。将肠癌细胞 HT29 在体外培养，并加入不同浓度（15 和 30mg/mL）的苍术内酯。然后通过细胞计数试剂盒-8（CCK8）检测细胞增殖活性，通过 EdU 染色检测阳性细胞的比例。通过酶联免疫吸附试验（ELISA）检测干扰素-γ（INF-γ）、肿瘤坏死因子-α（TNF-α）和基质金属蛋白酶-9（MMP-9）的含量，通过末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记（TUNEL）染色检测 HT29 细胞的凋亡。此外，通过逆转录-聚合酶链反应（RT-PCR）测定增殖、凋亡和 PI3K/AKT/mTOR 信号通路相关基因的信使核糖核酸（mRNA）水平，通过 Western 印迹分析 PI3K/AKT/mTOR 信号通路的表达。苍术内酯 15mg/mL 组细胞生长状态较差，密度较低，基本正常形态结构，苍术内酯 30mg/mL 组细胞数量明显减少，增殖活性明显减弱。苍术内酯 30mg/mL 组凋亡细胞明显增多。此外，苍术内酯 30mg/mL 组 INF-γ、TNF-α 和 MMP-9 均明显降低。苍术内酯 30mg/mL 组 B 细胞淋巴瘤-2（Bcl-2）、PI3K、AKT 和 mTOR 表达明显下降，NC 组升高，Caspase3 表达呈相反趋势。苍术内酯在适当浓度下通过抑制 PI3K/AKT/mTOR 信号通路抑制肠癌细胞增殖，促进其凋亡，可用于治疗结直肠癌等相关疾病。
