Yao Zhilin, Xu Xiaojuan, Huang Yinghong
Chengdu University of TCM, Chengdu 610075, China.
Saudi J Biol Sci. 2021 Dec;28(12):7077-7081. doi: 10.1016/j.sjbs.2021.08.011. Epub 2021 Aug 8.
Daidzin, 4', 7-dihydroxyisoflavone is an isoflavonic phytoestrogen present in leguminous plants. Traditional Chinese medicine utilizes daidzin to treat various diseases such diarrhea, fever, hepatitis, cardiac problems etc. In current study we examined the anticancer activity of daidzin against human cervical cancer in vitro. HeLa, human cervical cancer cell line was purchased from ATCC and the cells were cultured with DMEM medium. The cytotoxic effect of daidzin against HeLa cell line was analyzed with MTT assay. The IC-50 value was obtained at 20 µM hence the cells were treated with 20 µM of daidzin for further analysis. ROS generation was assessed with DCFH-DA staining and the induction of apoptosis was examined with Rhoadmine-123 staining. Acridine orange and ethidium bromide staining was done to examine the apoptotic and viable cells. Further the matrigel cell adhesion assay was done to analyze the inhibitory property of daidzin against cancer cell adhesion. Apoptotic induction of daidzin was examined by estimating the levels of Caspase 8 & 9 using ELISA technique. Inflammatory and cell proliferation signaling proteins were analyzed with qPCR analysis to confirm the anticancer activity of daidzin against human cervical cancer HeLa cell line. Daidzin significantly generated ROS and altered the mitochondrial membrane permeability in HeLa cell line. The results of AO/EtBr staining prove daidzin induced apoptosis in HeLa cell line and it also inhibited the cell adhesion property of HeLa which is reported in our matrigel cell adhesion assay. It also increased the caspases 8 & 9 which are key regulators of apoptosis. Daidzin significantly decreased the expression of inflammatory gene and cell proliferating signaling molecule. To, conclude our results confirm daidzin effectively decreased inflammation and induced apoptosis in human cervical cancer HeLa cell line.
大豆苷元，4'，7 - 二羟基异黄酮是一种存在于豆科植物中的异黄酮类植物雌激素。传统中医利用大豆苷元治疗各种疾病，如腹泻、发热、肝炎、心脏问题等。在当前研究中，我们在体外检测了大豆苷元对人宫颈癌的抗癌活性。人宫颈癌细胞系HeLa购自美国典型培养物保藏中心（ATCC），细胞用DMEM培养基培养。用MTT法分析大豆苷元对HeLa细胞系的细胞毒性作用。获得的半数抑制浓度（IC - 50）值为20 μM，因此用20 μM的大豆苷元处理细胞以进行进一步分析。用二氯荧光素二乙酸酯（DCFH - DA）染色评估活性氧（ROS）的产生，用罗丹明 - 123染色检测凋亡的诱导。进行吖啶橙和溴化乙锭染色以检测凋亡细胞和活细胞。此外，进行基质胶细胞黏附试验以分析大豆苷元对癌细胞黏附的抑制特性。通过酶联免疫吸附测定（ELISA）技术估计半胱天冬酶8和9水平来检测大豆苷元的凋亡诱导作用。用实时定量聚合酶链反应（qPCR）分析炎症和细胞增殖信号蛋白，以证实大豆苷元对人宫颈癌HeLa细胞系的抗癌活性。大豆苷元在HeLa细胞系中显著产生活性氧并改变线粒体膜通透性。吖啶橙/溴化乙锭（AO/EtBr）染色结果证明大豆苷元在HeLa细胞系中诱导凋亡，并且在我们的基质胶细胞黏附试验中它还抑制了HeLa细胞的黏附特性。它还增加了作为凋亡关键调节因子的半胱天冬酶8和9。大豆苷元显著降低炎症基因和细胞增殖信号分子的表达。总之，我们的结果证实大豆苷元有效减轻人宫颈癌HeLa细胞系中的炎症并诱导凋亡。
