Atractylodin may induce ferroptosis of human hepatocellular carcinoma cells

Abstract

Background: It has been reported that atractylodin has a potential antitumor effect. This study aimed to investigate the effects of atractylodin on Huh7 and Hccm hepatocellular carcinoma (HCC) cells and its molecular mechanism.Methods: Huh7 and Hccm cells were cultured in vitro, and their viability was detected by CCK-8 assay and the half inhibitory concentration (IC50) was calculated. The cells were treated with different concentrations of atractylodin, and the migration and invasion ability of cells was detected by scratch assay and Transwell assay. The cell cycle change and apoptosis rate were detected by flow cytometry. IlluminaHiSeq4000 platform was used for transcriptome sequencing, and the results were analyzed for gene differential expression, gene function, and signal pathway enrichment. Morphological changes of cells were detected by transmission electron microscopy, reactive oxygen species (ROS) levels were detected by DCFH-DA probe, and the expressions of ferroptosis related proteins GPX4, ACSL4, FTL, and TFR1 were detected by Western blot. Results:The results showed that atractylodin could inhibit the proliferation, migration, and invasion of Huh7 and Hccm cells, regulate the cell cycle, and induce cell apoptosis and G1 phase cell cycle arrest. In addition, it could significantly induce the increase of intracellular ROS levels, decrease the expression of GPX4 and FTL proteins, and up-regulate the expression of ACSL4 and TFR1 proteins.Conclusions: Atractylodin can inhibit the proliferation, migration, and invasion of Huh7 and Hccm liver cancer cells, and induce cell apoptosis and cell cycle arrest. In addition, our results suggest that atractylodin may induce ferroptosis in HCC cells by inhibiting the expression of GPX4 and FTL proteins, and upregulating the expression of ACSL4 and TFR1 proteins.

Integrated Analysis Reveals Critical Ferroptosis Regulators and FTL Contribute to Cancer Progression in Hepatocellular Carcinoma

“…As the main subunit of ferritin, FTL directly affects iron homeostasis ( Arber et al, 2016 ). Current studies have found that FTL may be regulated by the DNA damage response of serine/threonine kinase ATM, hypoxia, atractylodin, and other factors, resulting in ferroptosis ( Liu et al, 2020b ; Chen et al, 2020 ; He et al, 2021 ). FTL may also be involved in the development of tumors, such as the proliferation rate of HeLa cells and glioblastoma multiforme (GBM) cells ( Wu et al, 2016 ) and the drug resistance process of breast cancer ( Cozzi et al, 2004 ).…”

“…Circulating transcription of FTL is significantly upregulated in samples from HCC patients and maybe a new target for the diagnosis and treatment of HCC ( Wang et al, 2009 ; Sayeed et al, 2020 ). The decrease in FTL protein is associated with ferroptosis in HCC cells ( He et al, 2021 ), but its role and mechanism remain unknown.…”

Current Perspective of Traditional Chinese Medicines and Active Ingredients in the Therapy of Hepatocellular Carcinoma

“…59 In HCC cells, atractylodin was observed to induce the accumulation of lipid peroxides and ferroptosis by inhibiting GPX4 expression, leading to the suppression of HCC progression. 64 Therefore, it is suggested that ferroptosis plays a significant role in the inhibitory effect of atractylodin on HCC cells.…”

Perspectives and mechanisms for targeting ferroptosis in the treatment of hepatocellular carcinoma

“…It also promotes iron peroxidation and lipid ROS metabolism to induce ferroptosis in HCC cells ( Li Y et al, 2022 ). Atractylodin decreases GPX4 and FTL protein expression, upregulates ACSL4 and TFR1 protein expression, and increases ROS levels in liver cancer cells ( He et al, 2021 ). In recent years, several studies have reported that active components of Artemisia annua can induce ferroptosis and inhibit HCC.…”