荞麦壳黄酮诱导人肝癌HepG2细胞凋亡和抑制迁移的机制
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(1.吉林农业大学食品科学与工程学院 长春 130118;2.梧州学院食品与制药工程学院(六堡茶现代产业学院) 广西梧州 543002)

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吉林省科技发展计划项目(20200403159SF)


Mechanism of Buckwheat Hull Flavonoids in Inducement Apoptosis and Inhibiton Migration of Human Hepatocellular Carcinoma HepG2 Cells
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(1.College of Food Science and Engineering, Jilin Agricultural University, Changchun 130118;2.School of Food and Pharmaceutical Engineering (Guangxi Liupao Tea Modern Industry College), Wuzhou University, Wuzhou 543002, Guangxi)

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    摘要:

    探究荞麦壳黄酮对人肝癌HepG2细胞凋亡和迁移机制的影响。在用MTT比色法评价荞麦壳黄酮对人肺癌A549细胞、人宫颈癌Hela细胞、人肝癌HepG2细胞和人正常肝L-02细胞抑制效果的基础上,将HepG2细胞株作为试验对象,通过细胞形态学观察、Hoechst 33345/碘化丙啶(PI)双染、JC-1荧光探针检测、Annexin V-FITC/PI流式细胞术、伤口愈合试验和蛋白免疫印迹法,分析荞麦壳黄酮预处理对HepG2细胞凋亡和迁移的影响。结果表明:荞麦壳黄酮能够有效抑制A549细胞、Hela细胞和HepG2细胞的细胞活性,对HepG2细胞抑制效果最显著,50 μg/mL干预HepG2细胞48 h后,细胞存活率为53.62%,且对L-02细胞活性无影响。与未处理组相比,荞麦壳黄酮以剂量依赖性的方式抑制HepG2细胞增殖,降低线粒体膜电位,下调凋亡蛋白Bcl-2、Bcl-xl表达,上调FADD、cleaved-Caspase-3蛋白表达,并降低迁移蛋白MMP 9表达,且效果均显著高于阳性对照芦丁。研究结果说明荞麦壳黄酮具有抗肿瘤能力,其抑制HepG2细胞增殖可能与诱导线粒体、死亡受体细胞凋亡和抑制细胞迁移有关。

    Abstract:

    To investigate the effect of buckwheat hull flavonoids (BHFs) on apoptosis and migration of human hepatocellular carcinoma HepG2 cells. The inhibitory effect of BHFs on human lung cancer A549 cells, human cervical cancer Hela cells, human hepatocellular carcinoma HepG2 cells and human normal liver L-02 cells was evaluated by MTT colorimetric method, and HepG2 cells were further selected as experimental subjects. Cell morphology was analyzed by Hoechst 33345/PI double staining, JC-1 fluorescence probe detection, Annexin V-FITC/PI flow cytometry, wound healing assay, and Western blot analysis of BHFs pretreatment to investigate the effects of apoptosis and migration of HepG2 cells and the potential mechanism. The results showed that BHFs could effectively inhibit the viability of A549, Hela and HepG2 cells, the inhibitory effect on HepG2 cells was the most significant, the survival rate of HepG2 cells was 53.62% after treatment with 50 μg/mL for 48 h,and had no effect on L-02 cells viability. Compared with the untreated group, BHFs inhibited the proliferation of HepG2 cells in a dose-dependent manner, decreased mitochondrial membrane potential, down-regulated the expression of apoptotic proteins Bcl-2 and Bcl-XL, increased the expression of FADD and cleaved-Caspase-3, and decreased the expression of migratory protein MMP 9, and the effect was significantly superior to that of positive control of rutin. The results indicate that BHFs have anti-tumor ability, and the inhibition of HepG2 cell proliferation may be related to the induction of mitochondrial, death receptor cell apoptosis and the inhibition of cell migration.

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姜鑫,李天竹,王玥,刘子琦,王秀娟,朴春红.荞麦壳黄酮诱导人肝癌HepG2细胞凋亡和抑制迁移的机制[J].中国食品学报,2024,24(7):36-46

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  • 收稿日期:2023-07-04
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  • 在线发布日期: 2024-08-22
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