Liu Longfei, Qiu Yuan, Peng Zehao, Yu Zhongchao, Lu Hengzhe, Xie Rongjun, Mo Zhongcheng, Zhang Sen
Department of Colorectal & Anal Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China.
The Affiliated Nanhua Hospital, Department of Gastrointestinal Surgery, Hengyang Medical College, University of South China, Hengyang, Hunan 421001, China.
J Cancer. 2024 Nov 4;15(20):6741-6753. doi: 10.7150/jca.95775. eCollection 2024.
Colorectal cancer is among the most frequently diagnosed cancers with high mortality rates and poses a serious threat to human health. Genistein (Gen) has been found to have anti-colorectal cancer effects, however, the molecular mechanisms by which genistein elicits its effects on colorectal cancer (CRC) cells have not been fully elucidated. In this study, we investigated the oxidative state of colorectal cancer cells during the antitumor action of Genistein and whether it can exert its antitumor effects through ferroptosis. Current research on the oxidative state of Genistein indicates that it exhibits both antioxidant and pro-oxidant properties. Different drug concentrations were applied to colorectal cancer cells, after which cell viability and key markers of ferroptosis, including reactive oxygen species (ROS), malondialdehyde (MDA), and Fe, were measured. We found that genistein significantly reduced the viability of colorectal cancer cells, and the expression of ferroptosis markers increased in a concentration-dependent manner. Subsequently, we treated cells with the ferroptosis inhibitor fer-1 in combination with genistein and observed a partial reversal of ferroptosis markers. These findings suggest that genistein exerts its antitumor effect by promoting iron-dependent oxidative damage-induced ferroptosis. To further elucidate the mechanism underlying ferroptosis modulation, we examined the protein and mRNA expression levels of the classical key ferroptosis molecules SLC7A11 and GPX4. We found that the expression levels of these molecules decreased, with GPX4 exhibiting a greater decrease. Overexpression of GPX4 reversed the pro-ferroptotic effect of genistein, indicating that genistein promotes ferroptosis occurrence by downregulating GPX4 expression. When the drug was applied to colorectal cancer cells, the expression of the transcription factor FoxO3 increased. Treatment of cells with the FoxO3 inhibitor JY-2 in combination with other drugs resulted in antagonism of ferroptosis markers. These findings suggest that genistein induces ferroptosis in colorectal cancer cells through the FoxO3/SLC7A11/GPX4 signaling pathway, thereby inhibiting tumor growth.
结直肠癌是最常被诊断出的癌症之一，死亡率很高，对人类健康构成严重威胁。已发现染料木黄酮（Gen）具有抗结直肠癌作用，然而，染料木黄酮对结直肠癌细胞产生作用的分子机制尚未完全阐明。在本研究中，我们研究了染料木黄酮抗肿瘤作用过程中结直肠癌细胞的氧化状态，以及它是否能通过铁死亡发挥其抗肿瘤作用。目前关于染料木黄酮氧化状态的研究表明，它兼具抗氧化和促氧化特性。将不同药物浓度应用于结直肠癌细胞，然后测量细胞活力以及铁死亡的关键标志物，包括活性氧（ROS）、丙二醛（MDA）和铁。我们发现染料木黄酮显著降低了结直肠癌细胞的活力，并且铁死亡标志物的表达呈浓度依赖性增加。随后，我们用铁死亡抑制剂fer-1与染料木黄酮联合处理细胞，观察到铁死亡标志物有部分逆转。这些发现表明，染料木黄酮通过促进铁依赖性氧化损伤诱导的铁死亡发挥其抗肿瘤作用。为了进一步阐明铁死亡调节的潜在机制，我们检测了经典关键铁死亡分子SLC7A11和GPX4的蛋白质和mRNA表达水平。我们发现这些分子的表达水平下降，其中GPX4下降得更明显。GPX4的过表达逆转了染料木黄酮的促铁死亡作用，表明染料木黄酮通过下调GPX4表达促进铁死亡的发生。当将该药物应用于结直肠癌细胞时，转录因子FoxO3的表达增加。用FoxO3抑制剂JY-2与其他药物联合处理细胞导致铁死亡标志物的拮抗作用。这些发现表明，染料木黄酮通过FoxO3/SLC7A11/GPX4信号通路诱导结直肠癌细胞发生铁死亡，从而抑制肿瘤生长。
