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三氧化二砷对肝癌HepG2细胞迁移、侵袭及凋亡的影响

[Effects of arsenic trioxide on migration, invasion and apoptosis of hepatocellular carcinoma HepG2 cells].

作者信息

He Jia, Xu Bowen, Gao Wenbo, Su Guanyue, Yu Hongchi, Shen Yang, Liu Xiaoheng

机构信息

Institute of Biomedical Engineering, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu 610041, P.R.China.

National Engineering Research Center for Biomaterials, Chengdu 610065, P.R China.

出版信息

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2020 Feb 25;37(1):105-111. doi: 10.7507/1001-5515.201907044.

Abstract

The article aims to explore the optimal concentration of arsenic trioxide (As O ) on HepG2 of liver cancer cells, and the effect of As O on the migration, invasion and apoptosis of HepG2 cells. In this study, the activity of HepG2 cells treated with 0, 1, 2, 4, 8, 16, 32 μmol/L As O was tested by CCK-8 method, the semi-inhibitory concentration (IC50) was calculated, and the morphological changes of HepG2 cells were observed after the action of As O at IC50 concentration for 12, 24, 48 h. The effect of As O on cell migration and invasion ability was verified by wound healing experiment and Transwell invasion experiment. Western blot and qRT-PCR were used to detect the effects of As O on the gene and protein expression levels related to cell migration, invasion and apoptosis. The results showed that, compared with the control group, the activity of HepG2 cells decreased with the increase of the concentration of As O treatment, showing a dose-dependent effect, and its IC50 was 7.3 μmol/L. After 24 hours' treatment with 8 μmol/L As O , HepG2 cells underwent significant apoptosis, and its migration and invasion abilities were significantly reduced. In addition, the protein expression levels of RhoA, Cdc42, Rac1 and matrix metalloproteinase-9 (MMP-9) were down-regulated, the protein and mRNA expression levels of anti-apoptotic gene Bcl-2 were significantly down-regulated, and the protein and mRNA expression levels of pro-apoptotic genes Bax and Caspase-3 were significantly up-regulated. The above results indicate that certain concentration of As O can inhibit the migration and invasion of hepatocellular carcinoma cells and promote the apoptosis of hepatocellular carcinoma cells.

摘要

本文旨在探讨三氧化二砷(As₂O₃)对肝癌细胞HepG2的最佳浓度,以及As₂O₃对HepG2细胞迁移、侵袭和凋亡的影响。本研究采用CCK-8法检测0、1、2、4、8、16、32 μmol/L As₂O₃处理的HepG2细胞活性,计算半抑制浓度(IC50),并观察IC50浓度的As₂O₃作用12、24、48 h后HepG2细胞的形态变化。通过伤口愈合实验和Transwell侵袭实验验证As₂O₃对细胞迁移和侵袭能力的影响。采用Western blot和qRT-PCR检测As₂O₃对细胞迁移、侵袭和凋亡相关基因及蛋白表达水平的影响。结果显示,与对照组相比,HepG2细胞活性随As₂O₃处理浓度的增加而降低,呈剂量依赖性,其IC50为7.3 μmol/L。8 μmol/L As₂O₃处理24小时后,HepG2细胞发生明显凋亡,其迁移和侵袭能力显著降低。此外,RhoA、Cdc42、Rac1和基质金属蛋白酶-9(MMP-9)的蛋白表达水平下调,抗凋亡基因Bcl-2的蛋白和mRNA表达水平显著下调,促凋亡基因Bax和Caspase-3的蛋白和mRNA表达水平显著上调。上述结果表明,一定浓度的As₂O₃可抑制肝癌细胞的迁移和侵袭,促进肝癌细胞的凋亡。

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