Abstract
Scaphium affine 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 in vivo 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 in vitro and in vivo.
Highlights
Cancer is a disease caused by the random proliferation and metastasis of cells
HCT116 colorectal cancer cells showed % cell viability of 97.5%, 86.6%, 69.3%, 55.2%, and 41.2% at 50, 75, 100, 125, and 150 μg/ mL, respectively (Figure 2A), whereas HT29 colorectal cancer cells showed % cell viability of 100.3%, 93.5%, 82.2%, 69.8%, and 55.4%, respectively (Figure 2B). These results indicated the dosedependent cytotoxicity of Scaphium affine ethanol extracts (SAE) on HCT116 and HT29 colorectal cancer cells and that cytotoxicity was more effective on HCT116 colorectal cancer cells
The above changes were observed at 75 ug/ mL of low toxicity by stimulation. These results demonstrated that the concentration-dependent anoikisinducing effects of SAE in HCT116 colorectal cancer cells proceeds by inducing the intrinsic apoptotic pathway through the regulation of anoikis-related signaling proteins such as Epidermal growth factor receptor (EGFR), caveolin-1, Src, and Akt (Figure 4)
Summary
Cancer is a disease caused by the random proliferation and metastasis of cells. It is considered to be the leading cause of death in developed countries [1]. SAE Induce Anoikis in HCT116 drugs, colorectal diseases, metabolic syndrome, and heredity (only 5% of cases) are potential causes of colorectal cancer [3, 4]. Symptoms of colorectal cancer include alternating diarrhea and constipation as well as the appearance of mucus and blood in the feces. These symptoms can be accompanied by colon infarction, anemia, weight loss, and abdominal lumps [5, 6]
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