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解析红树莓诱导肝癌细胞凋亡的分子机制

Deciphering the Molecular Mechanism of Red Raspberry in Apoptosis of Liver Cancer Cells.

作者信息

Song Linlin, Li Qi, Shi Hui, Yue Hui

机构信息

College of Pharmacy, Jiamusi University, Jiamusi 154007, Heilongjiang, China.

Department of Biochemistry, Mudanjiang Medical University, Mudanjiang 157011, Heilongjiang, China.

出版信息

Evid Based Complement Alternat Med. 2022 Apr 27;2022:2026865. doi: 10.1155/2022/2026865. eCollection 2022.

Abstract

Red raspberry contains a variety of bioactive ingredients and has high edible and medicinal value. Red raspberry extractions (RREs) have strong antioxidant capacity and anticancer ability in vivo and in vitro. This study was to explore the specific mechanism of RREs inhibiting the proliferation of liver cancer HepG2 cells and provide a theoretical basis for the prevention and treatment of liver cancer by RREs. HepG2 cells were cultured in vitro, and MTT assay was adopted to detect the effect of RREs on HepG2 cell activity. Colony formation assay was applied to detect the growth and proliferation of cells, cell apoptosis was detected by flow cytometry, and dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay was adopted to detect the effect of RREs on the production of reactive oxygen species (ROS) in cells. The effect of RREs on cell mitochondrial membrane potential was evaluated by mitochondrial membrane potential assay kit with JC-1 (JC-1 assay), and western blot was used to detect the expression of apoptosis-related proteins (B-cell lymphoma-2 (Bcl-2), Bcl-2-associated (Bax), and Caspase-3), thus investigating the effect of RREs on the molecular mechanism of HepG2 cell apoptosis. The results showed that RREs could inhibit the proliferation activity of HepG2 cells and promote their apoptosis in a concentration-dependent manner. The level of ROS in HepG2 cells interfered by RREs increased markedly, while the cell mitochondrial membrane potential decreased sharply. As the concentration of HepG2 increased, the mitochondrial membrane potential reduced steeply. Western blot results showed that the expression of apoptosis-related protein Bcl-2 in the RREs treatment group dropped, but the expression of Bax and Caspase-3 rose. In summary, RREs could inhibit the proliferation of liver cancer HepG2 cells and promote their apoptosis. This inhibition might be executed by inducing HepG2 cells to produce ROS, a decrease in Bcl-2/Bax protein ratio, and an obvious reduction in mitochondrial membrane potential.

摘要

红树莓含有多种生物活性成分,具有较高的食用和药用价值。红树莓提取物(RREs)在体内外均具有较强的抗氧化能力和抗癌能力。本研究旨在探讨RREs抑制肝癌HepG2细胞增殖的具体机制,为RREs预防和治疗肝癌提供理论依据。体外培养HepG2细胞,采用MTT法检测RREs对HepG2细胞活性的影响。应用集落形成试验检测细胞的生长和增殖情况,通过流式细胞术检测细胞凋亡,采用二氯二氢荧光素二乙酸酯(DCFH-DA)试验检测RREs对细胞内活性氧(ROS)产生的影响。采用JC-1线粒体膜电位检测试剂盒(JC-1试验)评估RREs对细胞线粒体膜电位的影响,并用蛋白质免疫印迹法检测凋亡相关蛋白(B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关蛋白(Bax)和半胱天冬酶-3)的表达,从而研究RREs对HepG2细胞凋亡分子机制的影响。结果表明,RREs能够抑制HepG2细胞的增殖活性,并以浓度依赖的方式促进其凋亡。RREs干预后,HepG2细胞内ROS水平显著升高,而细胞线粒体膜电位急剧下降。随着RREs浓度的增加,线粒体膜电位急剧降低。蛋白质免疫印迹结果显示,RREs处理组凋亡相关蛋白Bcl-2的表达下降,但Bax和Caspase-3的表达升高。综上所述,RREs能够抑制肝癌HepG2细胞的增殖并促进其凋亡。这种抑制作用可能是通过诱导HepG2细胞产生ROS、降低Bcl-2/Bax蛋白比值以及显著降低线粒体膜电位来实现的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa9f/9068284/f9676d6b0e6f/ECAM2022-2026865.001.jpg

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