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硒代-β-乳球蛋白(Se-β-Lg)诱导 HepG2 细胞线粒体依赖性凋亡。

Seleno-β-lactoglobulin (Se-β-Lg) induces mitochondria-dependant apoptosis in HepG2 cells.

机构信息

Key Laboratory of Food Nutrition and Safety, Ministry of Education, College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, No. 29, 13th Street, TEDA, Tianjin, 300457, People's Republic of China.

QingYunTang Biotech (Beijing) Co., Ltd., Beijing, 100176, China.

出版信息

Mol Biol Rep. 2019 Oct;46(5):5025-5031. doi: 10.1007/s11033-019-04953-x. Epub 2019 Jul 30.

Abstract

Selenium compounds have been widely investigated as novel anticancer agents due to high efficacy and selectivity against cancer cells in recent years. This study aimed to research the potential inhibitory effects of seleno-β-lactoglobulin (Se-β-Lg) on HepG2 cells in vitro. MTT results demonstrated that the synthetized Se-β-Lg exhibited strong antitumor activity on HepG2 cells with few side effects on human normal cells (LO2) and relatively weaker cytotoxic effects compared to inorganic selenium (SeO). Scanning electron microscope (SEM), hoechst 33342/PI double staining, annexin V-FITC/PI staining and cell cycle detection results showed that Se-β-Lg could induce the apoptosis of HepG2 cells via arresting them in S and G2/M phases and lead to the obvious morphological changes (loss of adhesion, cell shrinkage, and membrane blebbing, membrane permeabilities and DNA fragmentation). Besides, JC-1 staining, western blotting (WB) and polymerase chain reaction (PCR) results showed that Se-β-Lg could gradually destroy the mitochondrial membrane potential of HepG2 cells, and finally resulting in the mitochondria-dependant apoptosis via up-regulation of Bax, Cytochrome c, Caspase-3 and down-regulation of Bcl-2. Our data could provide a theoretical basis for practical application of Se-β-Lg in food and drug industries.

摘要

近年来,由于硒化合物对癌细胞具有高效性和选择性,因此被广泛研究作为新型抗癌药物。本研究旨在研究合成的硒-β-乳球蛋白(Se-β-Lg)在体外对 HepG2 细胞的潜在抑制作用。MTT 结果表明,合成的 Se-β-Lg 对 HepG2 细胞具有很强的抗肿瘤活性,对人正常细胞(LO2)几乎没有副作用,且细胞毒性比无机硒(SeO)弱。扫描电子显微镜(SEM)、hoechst 33342/PI 双重染色、Annexin V-FITC/PI 染色和细胞周期检测结果表明,Se-β-Lg 可以通过将 HepG2 细胞阻滞在 S 和 G2/M 期来诱导细胞凋亡,并导致明显的形态变化(失去黏附、细胞收缩、细胞膜起泡、膜通透性和 DNA 片段化)。此外,JC-1 染色、western blotting(WB)和聚合酶链反应(PCR)结果表明,Se-β-Lg 可以逐渐破坏 HepG2 细胞的线粒体膜电位,最终通过上调 Bax、Cytochrome c、Caspase-3 和下调 Bcl-2 导致线粒体依赖性凋亡。我们的数据可为 Se-β-Lg 在食品和医药行业的实际应用提供理论基础。

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