He Yongfei, Fang Dalang, Liang Tianyi, Pang Hongbing, Nong Yingdan, Tang Libo, Yang Ziye, Lu Chunmiao, Han Xiao, Zhao Shuqi, Mo Shutian, Meng Yuhua, Han Chuangye, Peng Tao
Department of Hepatobiliary Surgery, the First Affiliated Hospital of Guangxi Medical University, Nanning, China.
Department of Breast and Thyroid Surgery, The Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, China.
Ann Transl Med. 2021 Oct;9(20):1535. doi: 10.21037/atm-21-4386.
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.
Huh7 and Hccm cells were cultured , 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.
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.
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 up-regulating the expression of ACSL4 and TFR1 proteins.
据报道，苍术素具有潜在的抗肿瘤作用。本研究旨在探讨苍术素对Huh7和Hccm肝癌细胞的影响及其分子机制。
培养Huh7和Hccm细胞，采用CCK-8法检测细胞活力并计算半数抑制浓度（IC50）。用不同浓度的苍术素处理细胞，采用划痕试验和Transwell试验检测细胞的迁移和侵袭能力。通过流式细胞术检测细胞周期变化和凋亡率。利用IlluminaHiSeq4000平台进行转录组测序，并对结果进行基因差异表达、基因功能和信号通路富集分析。通过透射电子显微镜检测细胞形态变化，用DCFH-DA探针检测活性氧（ROS）水平，并用蛋白质免疫印迹法检测铁死亡相关蛋白GPX4、ACSL4、FTL和TFR1的表达。
结果表明，苍术素可抑制Huh7和Hccm细胞的增殖、迁移和侵袭，调节细胞周期，诱导细胞凋亡并使细胞周期阻滞于G1期。此外，它可显著诱导细胞内ROS水平升高，降低GPX4和FTL蛋白的表达，并上调ACSL4和TFR1蛋白的表达。
苍术素可抑制Huh7和Hccm肝癌细胞的增殖、迁移和侵袭，并诱导细胞凋亡和细胞周期阻滞。此外，我们的结果表明，苍术素可能通过抑制GPX4和FTL蛋白的表达，上调ACSL4和TFR1蛋白的表达，从而诱导肝癌细胞发生铁死亡。
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