8-Nitrotryptanthrin inhibits colorectal cancer progression via TGF-β/SMAD and PI3K/AKT/mTOR pathways

ObjectiveTo investigate the anticancer effects and underlying mechanisms of 8-nitrotryptanthrin (8-Nitrotryp) against colorectal cancer (CRC).MethodsThe effects of 8-Nitrotryp on proliferation, colony formation, and migration were evaluated in HCT116 and SW480 cells, with comparisons to its parent c...

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Main Authors: Cheng-Yu Sun, Kai-Ping Cong, Dan-Dan Zhao, En-Guo Fan, Ming-Quan Guo, Zheng-Guo Zhang
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-08-01
Series:Frontiers in Pharmacology
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Online Access:https://www.frontiersin.org/articles/10.3389/fphar.2025.1576673/full
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author Cheng-Yu Sun
Kai-Ping Cong
Kai-Ping Cong
Dan-Dan Zhao
En-Guo Fan
Ming-Quan Guo
Ming-Quan Guo
Zheng-Guo Zhang
author_facet Cheng-Yu Sun
Kai-Ping Cong
Kai-Ping Cong
Dan-Dan Zhao
En-Guo Fan
Ming-Quan Guo
Ming-Quan Guo
Zheng-Guo Zhang
author_sort Cheng-Yu Sun
collection DOAJ
description ObjectiveTo investigate the anticancer effects and underlying mechanisms of 8-nitrotryptanthrin (8-Nitrotryp) against colorectal cancer (CRC).MethodsThe effects of 8-Nitrotryp on proliferation, colony formation, and migration were evaluated in HCT116 and SW480 cells, with comparisons to its parent compound tryptanthrin (Tryp). Mitochondrial membrane potential (MMP) was assessed using JC-1 staining, and early apoptosis was analyzed by flow cytometry. Proteomic analysis and Western blotting were employed to examine the modulation of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mechanistic target of the rapamycin (mTOR) pathway and transforming growth factor-β (TGF-β)/Sma- and Mad-related proteins (SMAD) signaling pathways, as well as epithelial–mesenchymal transition (EMT).Results8-Nitrotryp significantly inhibited proliferation of HCT116 (IC50 = 0.81–1.08 μM; P < 0.001) and SW480 cells (IC50 = 0.76–1.59 μM; P < 0.001), suppressed colony formation of HCT116 (P < 0.001 at 1 μM) and SW480 cells (P < 0.001 at 2 μM), and inhibited migration in a dose-dependent manner (0.5–2 μM), demonstrating greater potency than Tryp. It also suppressed MMP and induced early apoptosis in HCT116 (P < 0.001 at 1 μM) and SW480 cells (P < 0.001 at 0.5 μM). Proteomic analysis and Western blotting revealed that 8-Nitrotryp downregulated PI3K expression, inhibited the phosphorylation of AKT and mTOR, and reduced TGF-β1-induced SMAD2 phosphorylation. Additionally, 8-Nitrotryp suppressed the EMT process.Conclusion8-Nitrotryp inhibits CRC progression by modulating the TGF-β/SMAD and PI3K/AKT/mTOR pathways, highlighting its potential as a multi-target therapeutic agent for CRC and warranting its further investigation.Novelty and ImpactCRC is a global health challenge with limited treatments for advanced stages. This study provides the first evidence of 8-Nitrotryp’s antitumor efficacy in CRC, demonstrating its dual inhibitory activity on the TGF-β/SMAD and PI3K/AKT/mTOR pathways. Compared to Tryp, 8-Nitrotryp exhibits markedly enhanced potency, with lower IC50 values due to the introduction of a nitro group. Furthermore, the suppression of EMT is mechanistically linked to TGF-β/SMAD pathway inhibition. These findings suggest 8-Nitrotryp’s potential as a novel therapeutic for CRC.
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spelling doaj-art-a378ed7a57e0460284aa876cb1918dbd2025-08-22T05:26:53ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122025-08-011610.3389/fphar.2025.157667315766738-Nitrotryptanthrin inhibits colorectal cancer progression via TGF-β/SMAD and PI3K/AKT/mTOR pathwaysCheng-Yu Sun0Kai-Ping Cong1Kai-Ping Cong2Dan-Dan Zhao3En-Guo Fan4Ming-Quan Guo5Ming-Quan Guo6Zheng-Guo Zhang7Department of Colorectal Surgery, The Affiliated Xuzhou Clinical College of Xuzhou Medical University, Xuzhou, Jiangsu, ChinaLaboratory of Advanced Theranostic Materials and Technology, Ningbo Institute of Materials Technology and Engineering, Chinese Academy of Sciences, Ningbo, ChinaNingbo Key Laboratory of Biomedical Imaging Probe Materials and Technology, Zhejiang International Cooperation Base of Biomedical Materials Technology and Application, Ningbo Cixi Institute of Biomedical Engineering, Ningbo, ChinaSchool of Life Science, Jiangsu Normal University, Xuzhou, Jiangsu, ChinaPeking Union Medical College, Beijing, ChinaLaboratory of Advanced Theranostic Materials and Technology, Ningbo Institute of Materials Technology and Engineering, Chinese Academy of Sciences, Ningbo, ChinaNingbo Key Laboratory of Biomedical Imaging Probe Materials and Technology, Zhejiang International Cooperation Base of Biomedical Materials Technology and Application, Ningbo Cixi Institute of Biomedical Engineering, Ningbo, ChinaDepartment of Colorectal Surgery, The Affiliated Xuzhou Clinical College of Xuzhou Medical University, Xuzhou, Jiangsu, ChinaObjectiveTo investigate the anticancer effects and underlying mechanisms of 8-nitrotryptanthrin (8-Nitrotryp) against colorectal cancer (CRC).MethodsThe effects of 8-Nitrotryp on proliferation, colony formation, and migration were evaluated in HCT116 and SW480 cells, with comparisons to its parent compound tryptanthrin (Tryp). Mitochondrial membrane potential (MMP) was assessed using JC-1 staining, and early apoptosis was analyzed by flow cytometry. Proteomic analysis and Western blotting were employed to examine the modulation of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mechanistic target of the rapamycin (mTOR) pathway and transforming growth factor-β (TGF-β)/Sma- and Mad-related proteins (SMAD) signaling pathways, as well as epithelial–mesenchymal transition (EMT).Results8-Nitrotryp significantly inhibited proliferation of HCT116 (IC50 = 0.81–1.08 μM; P < 0.001) and SW480 cells (IC50 = 0.76–1.59 μM; P < 0.001), suppressed colony formation of HCT116 (P < 0.001 at 1 μM) and SW480 cells (P < 0.001 at 2 μM), and inhibited migration in a dose-dependent manner (0.5–2 μM), demonstrating greater potency than Tryp. It also suppressed MMP and induced early apoptosis in HCT116 (P < 0.001 at 1 μM) and SW480 cells (P < 0.001 at 0.5 μM). Proteomic analysis and Western blotting revealed that 8-Nitrotryp downregulated PI3K expression, inhibited the phosphorylation of AKT and mTOR, and reduced TGF-β1-induced SMAD2 phosphorylation. Additionally, 8-Nitrotryp suppressed the EMT process.Conclusion8-Nitrotryp inhibits CRC progression by modulating the TGF-β/SMAD and PI3K/AKT/mTOR pathways, highlighting its potential as a multi-target therapeutic agent for CRC and warranting its further investigation.Novelty and ImpactCRC is a global health challenge with limited treatments for advanced stages. This study provides the first evidence of 8-Nitrotryp’s antitumor efficacy in CRC, demonstrating its dual inhibitory activity on the TGF-β/SMAD and PI3K/AKT/mTOR pathways. Compared to Tryp, 8-Nitrotryp exhibits markedly enhanced potency, with lower IC50 values due to the introduction of a nitro group. Furthermore, the suppression of EMT is mechanistically linked to TGF-β/SMAD pathway inhibition. These findings suggest 8-Nitrotryp’s potential as a novel therapeutic for CRC.https://www.frontiersin.org/articles/10.3389/fphar.2025.1576673/fullcolorectal neoplasms8-nitrotryptanthrinsignal transductiontransforming growth factor betaphosphatidylinositol 3-kinases
spellingShingle Cheng-Yu Sun
Kai-Ping Cong
Kai-Ping Cong
Dan-Dan Zhao
En-Guo Fan
Ming-Quan Guo
Ming-Quan Guo
Zheng-Guo Zhang
8-Nitrotryptanthrin inhibits colorectal cancer progression via TGF-β/SMAD and PI3K/AKT/mTOR pathways
Frontiers in Pharmacology
colorectal neoplasms
8-nitrotryptanthrin
signal transduction
transforming growth factor beta
phosphatidylinositol 3-kinases
title 8-Nitrotryptanthrin inhibits colorectal cancer progression via TGF-β/SMAD and PI3K/AKT/mTOR pathways
title_full 8-Nitrotryptanthrin inhibits colorectal cancer progression via TGF-β/SMAD and PI3K/AKT/mTOR pathways
title_fullStr 8-Nitrotryptanthrin inhibits colorectal cancer progression via TGF-β/SMAD and PI3K/AKT/mTOR pathways
title_full_unstemmed 8-Nitrotryptanthrin inhibits colorectal cancer progression via TGF-β/SMAD and PI3K/AKT/mTOR pathways
title_short 8-Nitrotryptanthrin inhibits colorectal cancer progression via TGF-β/SMAD and PI3K/AKT/mTOR pathways
title_sort 8 nitrotryptanthrin inhibits colorectal cancer progression via tgf β smad and pi3k akt mtor pathways
topic colorectal neoplasms
8-nitrotryptanthrin
signal transduction
transforming growth factor beta
phosphatidylinositol 3-kinases
url https://www.frontiersin.org/articles/10.3389/fphar.2025.1576673/full
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