Kawk Hye Won, Nam Gun-He, Kim Myeong Jin, Kim Sang-Yong, Kim Young-Min
Department of Biological Science and Biotechnology, College of Life Science and Nano Technology, Hannam University, Daejeon, South Korea.
Department of Food Science and Bio Technology, Shinansan University, Ansan, South Korea.
Front Oncol. 2021 May 20;11:621346. doi: 10.3389/fonc.2021.621346. eCollection 2021.
ethanol extracts (SAE) is a species that has been shown to contain various physiological effects; however, its anticancer effects have yet to be revealed. We qualitatively evaluated β-sitosterol in SAE through high-performance liquid chromatography (HPLC). The cytotoxicity in HCT116 and HT29 colorectal cancer cells and CCD841 normal colon cells was confirmed through WST-1 assays. Selective cytotoxicity was observed in colorectal cancer cells, with greater cytotoxicity demonstrated in the HCT116 cell line. As such, the HCT116 colorectal cell line was selected for subsequent experiments. After HCT116 cells were treated with SAE, it was confirmed that the apoptosis rate was increased in a SAE dose-dependent manner through Annexin V assay. SAE further showed dose-dependent suppression of invasion through invasion assays. Anoikis induction through the EGFR/Akt pathway in HCT116 colorectal cancer cells was confirmed by Western blotting. The tumor suppressive effects of SAE was assessed using a xenograft model of human HCT116 colorectal cancer cells. As a result, we confirmed that SAE decreased tumor size in a dose-dependent manner and that p-EGFR and cleaved-caspase 3 in tumors were also regulated in a dose-dependent manner. This study showed that SAE, by containing β-sitosterol with proven anticancer effects, induces anoikis through the EGFR/Akt pathway in HCT116 colorectal cancer cells both and .
乙醇提取物（SAE）是一种已被证明具有多种生理效应的物质；然而，其抗癌作用尚未明确。我们通过高效液相色谱法（HPLC）对SAE中的β-谷甾醇进行了定性评估。通过WST-1检测法确认了SAE对HCT116和HT29结肠癌细胞以及CCD841正常结肠细胞的细胞毒性。在结肠癌细胞中观察到了选择性细胞毒性，HCT116细胞系表现出更大的细胞毒性。因此，选择HCT116结肠癌细胞系进行后续实验。用SAE处理HCT116细胞后，通过膜联蛋白V检测法确认细胞凋亡率以SAE剂量依赖性方式增加。通过侵袭实验，SAE进一步显示出剂量依赖性的侵袭抑制作用。通过蛋白质印迹法证实了HCT116结肠癌细胞中通过EGFR/Akt途径诱导失巢凋亡。使用人HCT116结肠癌细胞的异种移植模型评估了SAE的肿瘤抑制作用。结果，我们证实SAE以剂量依赖性方式减小肿瘤大小，并且肿瘤中的p-EGFR和裂解的半胱天冬酶3也以剂量依赖性方式受到调节。这项研究表明，SAE通过含有具有已证实抗癌作用的β-谷甾醇，在HCT116结肠癌细胞中通过EGFR/Akt途径诱导失巢凋亡。
