三氧化二砷对肝癌细胞HepG2迁移、侵袭和凋亡的影响

本文旨在探究三氧化二砷(As2O3)对肝癌细胞HepG2作用的最佳浓度,以及此浓度As2O3对HepG2细胞迁移、侵袭和凋亡的影响。本研究采用CCK-8法检验0、1、2、4、8、16、32μmol/L As2O3处理后HepG2细胞活力,计算半抑制浓度(IC50),并且观察IC50浓度As2O3作用12、24、48 h后HepG2细胞形态变化。通过伤口愈合实验和Transwell侵袭实验验证As2O3对细胞迁移侵袭能力的影响。Western blot和qRT-PCR实验检测As2O3对细胞迁移、侵袭及凋亡相关蛋白和基因表达水平的影响。结果显示,与对照组相比,HepG2细胞活力随As2O3处理浓度的升高而降低,呈现剂量依懒性,其IC50为7.3μmol/L;8μmol/L As2O3处理24 h后,HepG2细胞发生明显凋亡,且其迁移和侵袭能力显著降低;此外,细胞中RhoA、Cdc42、Rac1、基质金属蛋白酶-9(MMP-9)蛋白表达水平下调,抑凋亡基因Bcl-2的蛋白和mRNA表达水平显著下调,而促凋亡基因Bax、Caspase-3的蛋白和mRNA表达水平显著上调。以上结果说明一定浓度的As2O3能够抑制肝癌细胞迁移和侵袭,促进肝癌细胞凋亡。

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

The article aims to explore the optimal concentration of arsenic trioxide(As2O3)on HepG2 of liver cancer cells,and the effect of As2O3 on the migration,invasion and apoptosis of HepG2 cells.In this study,the activity of HepG2 cells treated with 0,1,2,4,8,16,32μmol/L As2O3 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 As2O3 at IC50 concentration for 12,24,48 h.The effect of As2O3 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 As2O3 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 As2O3 treatment,showing a dose-dependent effect,and its IC50 was 7.3μmol/L.After 24 hours’treatment with 8μmol/L As2O3,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 As2O3 can inhibit the migration and invasion of hepatocellular carcinoma cells and promote the apoptosis of hepatocellular carcinoma cells.