Department of Biological Sciences, Gachon University, Seongnam, South Korea.
Department of Bio and Chemical Engineering, Hongik University, Sejong 30016, South Korea.
Food Chem Toxicol. 2018 Jun;116(Pt B):315-322. doi: 10.1016/j.fct.2018.04.053. Epub 2018 Apr 24.
The flavonoid myricetin (MYR) is derived from vegetables and fruits. It has been shown to exert anti-cancer effects in various cell lines; however, the exact mechanism underlying these effects is yet to be elucidated. In this study, we evaluated the anti-cancer effects induced by MYR treatment in colon cancer HCT-15 cells. To detect cell proliferation, we conducted MTT assay and real time-cell electronic sensing (RT-CES). We next performed comet assay and Annexin V and PI staining to detect cellular apoptotic features. After that, we conducted two-dimensional electrophoresis (2-DE) analysis to identify apoptotic proteins. The results of this analysis revealed that eight spots were differentially expressed. Among the spots, we selected nucleoside diphosphate kinase (NDPK) for further investigation, as it has been shown to regulate cancer cell apoptosis and metastasis. After that, we conducted realtime-PCR and western blot to detect the expression of NDPK mRNA and protein and wound-healing assay to detect cell migration and invasion. Finally, we performed NDPK siRNA transfection study and the results showed that NDPK knockdown inhibited apoptosis. Based on these collective results, we suggest that MYR induces apoptosis in human colon cancer HCT-15 cells selectively by increasing the expression of NDPK and other caspase-regulated apoptosis proteins.
杨梅素（MYR）是一种从蔬菜和水果中提取的类黄酮，已被证实对多种细胞系具有抗癌作用；然而，其确切的作用机制仍有待阐明。在本研究中，我们评估了 MYR 处理对结肠癌细胞 HCT-15 诱导的抗癌作用。为了检测细胞增殖，我们进行了 MTT 检测和实时细胞电子感应（RT-CES）实验。接下来，我们进行彗星实验和 Annexin V 和 PI 染色，以检测细胞凋亡特征。之后，我们进行二维电泳（2-DE）分析，以鉴定凋亡蛋白。该分析的结果显示，有 8 个点差异表达。在这些点中，我们选择核苷二磷酸激酶（NDPK）进行进一步研究，因为它已被证明可以调节癌细胞凋亡和转移。之后，我们进行实时 PCR 和 Western blot 检测 NDPK mRNA 和蛋白的表达，并进行划痕愈合实验检测细胞迁移和侵袭。最后，我们进行了 NDPK siRNA 转染研究，结果表明 NDPK 敲低抑制了细胞凋亡。基于这些综合结果，我们认为 MYR 通过增加 NDPK 和其他半胱天冬酶调控的凋亡蛋白的表达，选择性地诱导人结肠癌细胞 HCT-15 的凋亡。
