Wu Yingchao, Pi Dajin, Chen Yiliu, Zuo Qian, Zhou Shuyao, Ouyang Mingzi
School of Traditional Chinese Medicine, Jinan University, Guangzhou, Guangdong 510632, China.
MOE Key Laboratory of Tumor Molecular Biology and Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, College of Life Science and Technology, Jinan University, Guangzhou, Guangdong 510632, China.
Evid Based Complement Alternat Med. 2022 May 29;2022:6177553. doi: 10.1155/2022/6177553. eCollection 2022.
Colorectal cancer (CRC) is a severe threat to human health. Ginsenosides such as ginsenoside Rh4 have been widely studied in the antitumor field. Here, we investigated the antiproliferative activity and mechanism of Rh4 against CRC in vivo and in vitro. The CRC xenograft model showed that Rh4 inhibited xenograft tumor growth with few side effects ( < 0.05). As determined by MTT colorimetric assays, Western blotting, and immunohistochemical analysis, Rh4 effectively inhibited CRC cell proliferation through autophagy and ferroptosis ( < 0.05). Rh4 significantly upregulated autophagy and ferroptosis marker expression in CRC cells and xenograft tumor tissues in the present study ( < 0.05). Interestingly, the ferroptosis inhibitor ferrostatin-1 (Fer-1) reversed Rh4-induced ferroptosis ( < 0.05). Moreover, the autophagy inhibitor 3-methyladenine (3-MA) also reversed Rh4-induced ferroptosis ( < 0.05). These results indicate that Rh4-induced ferroptosis is regulated via the autophagy pathway. In addition, Rh4 increased reactive oxygen species (ROS) accumulation, leading to the activation of the ROS/p53 signaling pathway ( < 0.05). Transcriptome sequencing also confirmed this ( < 0.05). Moreover, the ROS scavenger N-acetyl-cysteine (NAC) reversed the inhibitory effect of Rh4 on CRC cells ( < 0.05). Therefore, this study proves that Rh4 inhibits cancer cell proliferation by activating the ROS/p53 signaling pathway and activating autophagy to induce ferroptosis, which provides necessary scientific evidence of the great anticancer potential of Rh4.
结直肠癌（CRC）对人类健康构成严重威胁。人参皂苷如人参皂苷Rh4在抗肿瘤领域已得到广泛研究。在此，我们研究了Rh4在体内和体外对CRC的抗增殖活性及机制。CRC异种移植模型表明，Rh4抑制异种移植肿瘤生长，且副作用较小（<0.05）。通过MTT比色法、蛋白质印迹法和免疫组织化学分析确定，Rh4通过自噬和铁死亡有效抑制CRC细胞增殖（<0.05）。在本研究中，Rh4显著上调CRC细胞和异种移植肿瘤组织中自噬和铁死亡标志物的表达（<0.05）。有趣的是，铁死亡抑制剂铁抑素-1（Fer-1）逆转了Rh4诱导的铁死亡（<0.05）。此外，自噬抑制剂3-甲基腺嘌呤（3-MA）也逆转了Rh4诱导的铁死亡（<0.05）。这些结果表明，Rh4诱导的铁死亡是通过自噬途径调节的。此外，Rh4增加了活性氧（ROS）的积累，导致ROS/p53信号通路的激活（<0.05）。转录组测序也证实了这一点（<0.05）。此外，ROS清除剂N-乙酰半胱氨酸（NAC）逆转了Rh4对CRC细胞的抑制作用（<0.05）。因此，本研究证明Rh4通过激活ROS/p53信号通路和激活自噬以诱导铁死亡来抑制癌细胞增殖，这为Rh4具有巨大抗癌潜力提供了必要的科学证据。
