Department of Urology, The First Affiliated Hospital of Harbin Medical University, 23 Postal Street, Harbin, 150000, Heilongjiang, China.
Key Laboratory of Hepatosplenic Surgery, Ministry of Education, The First Affiliated Hospital of Harbin Medical University, Harbin, 150000, Heilongjiang, China.
Sci Rep. 2024 Jan 10;14(1):1009. doi: 10.1038/s41598-023-50476-8.
Patients with bladder cancer (BLCA) still show high recurrence after surgery and chemotherapy. Hesperetin (HE), as a natural compound, has attracted researchers' attention due to its low toxicity and easy access. However, the inhibitory effect of HE on BLCA remains unknown. The hub genes and enrichment pathways regulated by HE in the treatment of BLCA were predicted by network pharmacology. The molecular docking of HE and hub proteins was visualized. Colony and CCK8 assays were used to test cell proliferation, and BLCA migration was confirmed by transwell and wound healing assays. In addition, the occurrence of apoptosis and ferroptosis was demonstrated by Hoechst staining, transmission electron microscopy (TEM) and ROS (reactive oxygen species) assay. Western Blotting was performed to validate the hub proteins, target functions and pathways. SRC, PIK3R1 and MAPK1 were identified as hub targets for HE in BLCA, involving the PI3k/AKT pathway. Furthermore, HE inhibited the proliferation and migration of BLCA cells. The MMP2/MMP9 proteins were significantly inhibited by HE. The increased expression of Bax and cleaved caspase-3 indicated that HE could promote BLCA cell apoptosis. In addition, Hoechst staining revealed concentrated and illuminated apoptotic nuclei. The activation of ROS and the decline of GPX4 expression suggested that HE might induce ferroptosis as an anti-BLCA process. Shrunk mitochondria and apoptotic bodies were observed in BLCA cells treated with HE, with reduced or absent mitochondrial cristae. We propose for the first time that HE could inhibit the proliferation and migration of BLCA cells and promote apoptosis and ferroptosis. HE may act by targeting proteins such as SRC, PIK3R1 and MAPK1 and the PI3K/AKT pathway.
患有膀胱癌 (BLCA) 的患者在手术后和化疗后仍表现出高复发率。橙皮素 (HE) 作为一种天然化合物，由于其低毒性和易得性，引起了研究人员的关注。然而，HE 对 BLCA 的抑制作用尚不清楚。通过网络药理学预测 HE 治疗 BLCA 调节的枢纽基因和富集途径。可视化 HE 和枢纽蛋白的分子对接。使用集落和 CCK8 测定法测试细胞增殖，通过 Transwell 和划痕愈合测定法证实 BLCA 迁移。此外，通过 Hoechst 染色、透射电子显微镜 (TEM) 和 ROS (活性氧) 测定法证明细胞凋亡和铁死亡的发生。通过 Western Blotting 验证枢纽蛋白、靶功能和途径。SRC、PIK3R1 和 MAPK1 被确定为 HE 在 BLCA 中的枢纽靶标，涉及 PI3k/AKT 途径。此外，HE 抑制 BLCA 细胞的增殖和迁移。HE 显著抑制 MMP2/MMP9 蛋白。Bax 和 cleaved caspase-3 的表达增加表明 HE 可以促进 BLCA 细胞凋亡。此外，Hoechst 染色显示凋亡核浓缩和发光。ROS 的激活和 GPX4 表达的下降表明 HE 可能作为抗 BLCA 过程诱导铁死亡。用 HE 处理的 BLCA 细胞中观察到线粒体收缩和凋亡小体，线粒体嵴减少或消失。我们首次提出，HE 可以抑制 BLCA 细胞的增殖和迁移，促进细胞凋亡和铁死亡。HE 可能通过靶向 SRC、PIK3R1 和 MAPK1 等蛋白以及 PI3K/AKT 途径发挥作用。
