橙皮苷通過網絡藥理學和分子對接，通過PI3K/AKT通路促進膀胱癌細胞死亡

2024年1月，1.郵政街23號哈爾濱醫科大學**附屬醫院泌尿外科；2.哈爾濱醫科大學**附屬醫院肝脾外科教育部重點實驗室；3.南昌大學**附屬醫院超聲醫學科；4.海南醫科大學附屬醫院海南總醫院神經外科;（1. Department of Urology, The First Afliated Hospital of Harbin Medical University, 23 Postal Street,2. Key Laboratory of Hepatosplenic Surgery, Ministry of Education, The First Afliated Hospital of Harbin Medical University,3. Department of Ultrasound Medicine, The First Afliated Hospital of Nanchang University,4. Department of Neurosurgery, Hainan General Hospital, Hainan Afliated Hospital of Hainan Medical University,）Youcheng Xiu1 & Zan Liu1研究團隊在《Scientific Reports》上發表論文：

“Hesperetin promotes bladder cancer cells death via the PI3K/AKT pathway by network pharmacology and molecular docking”

“橙皮苷通過網絡藥理學和分子對接，通過PI3K/AKT通路促進膀胱癌細胞死亡”

Abstract：

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 efect 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 confrmed 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 identifed 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 signifcantly 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 frst 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.

摘要：

膀胱癌(BLCA)患者在手術和化療后仍有高復發率。橙皮苷(Hesperetin, HE)作為一種天然化合物，因其毒性低、易于獲取而備受關注。然而，HE對BLCA的抑制作用尚不清楚。利用網絡藥理學方法預測HE調控的中樞基因和富集途徑在BLCA治療中的作用。可視化了HE蛋白與樞紐蛋白的分子對接。用菌落和CCK8檢測細胞增殖，用transwell和傷口愈合檢測證實BLCA遷移。此外，通過Hoechst染色、透射電鏡(TEM)和活性氧(ROS)測定證實了細胞凋亡和鐵下垂的發生。Western Blotting驗證中心蛋白、靶功能和通路。SRC, PIK3R1和MAPK1被確定為BLCA中HE的樞紐靶點，涉及PI3k/AKT通路。此外，HE抑制BLCA細胞的增殖和遷移。HE顯著抑制MMP2/MMP9蛋白表達。Bax和cleaved caspase-3的表達增加表明HE可促進BLCA細胞凋亡。此外，Hoechst染色顯示凋亡核的集中和照亮。ROS的激活和GPX4表達的下降提示HE可能通過抗blca過程誘導鐵下垂。HE處理的BLCA細胞線粒體縮小，凋亡小體出現，線粒體嵴減少或缺失。我們**提出HE可以抑制BLCA細胞的增殖和遷移，促進細胞凋亡和鐵下垂。HE可能通過靶向SRC、PIK3R1和MAPK1等蛋白以及PI3K/AKT通路發揮作用。

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