Aniline TFPA enhances camptothecin-induced anti-NSCLC by modulating oxidative stress and impairing autophagy

Abstract Background Camptothecin (CPT) derivatives are widely used in cancer therapies, but their efficacy can be attenuated by resistance mechanisms such as autophagy. We recently showed that the aniline compound 4-[4-(4-aminophenoxy)-2,3,5,6-tetrafluorophenoxy] aniline (TFPA) can potently increase...

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Main Authors: Han-Lin Chou, I-Ling Lin, Yei-Tsung Chen, Wen-Tsan Chang, Ann Yu, Wei-Chun Chen, Chang-Yi Wu, Shean-Jaw Chiou, Chih-Wen Shu, Chien-Chih Chiu, Pei-Feng Liu
Format: Article
Language:English
Published: BMC 2025-03-01
Series:Cancer Cell International
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Online Access:https://doi.org/10.1186/s12935-025-03657-6
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author Han-Lin Chou
I-Ling Lin
Yei-Tsung Chen
Wen-Tsan Chang
Ann Yu
Wei-Chun Chen
Chang-Yi Wu
Shean-Jaw Chiou
Chih-Wen Shu
Chien-Chih Chiu
Pei-Feng Liu
author_facet Han-Lin Chou
I-Ling Lin
Yei-Tsung Chen
Wen-Tsan Chang
Ann Yu
Wei-Chun Chen
Chang-Yi Wu
Shean-Jaw Chiou
Chih-Wen Shu
Chien-Chih Chiu
Pei-Feng Liu
author_sort Han-Lin Chou
collection DOAJ
description Abstract Background Camptothecin (CPT) derivatives are widely used in cancer therapies, but their efficacy can be attenuated by resistance mechanisms such as autophagy. We recently showed that the aniline compound 4-[4-(4-aminophenoxy)-2,3,5,6-tetrafluorophenoxy] aniline (TFPA) can potently increase CPT cytotoxicity against non-small cell lung cancer (NSCLC) cells. The purpose of this study was to evaluate whether TFPA improves CPT-based chemotherapy by modulating autophagy and other cell death pathways in NSCLC models. Methods Two NSCLC cell lines, A549 and H1299, were tested. The synergism of CPT and TFPA was evaluated by trypan blue exclusion and colony formation assays. Annexin V staining was used for the detection of apoptosis, and autophagy was assessed by acridine orange staining and immunofluorescence. Flow cytometry-based dihydroethidium staining was used to assess oxidative stress. Changes in the expression of apoptosis-associated factors and autophagy-associated factors were determined by Western blot assays. The synergism of CPT and TFPA was validated using a zebrafish xenograft assay. Results The accumulation of markers for lysosomal expansion (LAMP2) and degradation (cathepsin D) and markers for autophagosome formation (LC3B-II) suggested that blockage of autolysosome formation might impair autophagy in CPT-treated NSCLC cells and subsequently lead to autophagic cell death. Cotreatment with TFPA and CPT induced cell death by increasing the production of reactive oxygen species, which contributed to autophagic impairment and eventually apoptotic cell death in NSCLC cells. Conclusions Our present work suggests that increased autophagic impairment induced by the combination of CPT and TFPA contributes to the apoptotic cell death of lung cancer cells.
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spelling doaj-art-d2a45c8cf10f41e3b0e867a13039d76e2025-08-20T02:47:07ZengBMCCancer Cell International1475-28672025-03-0125111510.1186/s12935-025-03657-6Aniline TFPA enhances camptothecin-induced anti-NSCLC by modulating oxidative stress and impairing autophagyHan-Lin Chou0I-Ling Lin1Yei-Tsung Chen2Wen-Tsan Chang3Ann Yu4Wei-Chun Chen5Chang-Yi Wu6Shean-Jaw Chiou7Chih-Wen Shu8Chien-Chih Chiu9Pei-Feng Liu10Department of Biotechnology, Kaohsiung Medical UniversityDepartment of Medical Laboratory Science and Biotechnology, Kaohsiung Medical UniversityDepartment of Life Sciences, Institute of Genome Sciences, National Yang Ming Chiao Tung UniversityDivision of General and Digestive Surgery, Department of Surgery, Kaohsiung Medical University HospitalDepartment of Biotechnology, Kaohsiung Medical UniversityDepartment of Biotechnology, Kaohsiung Medical UniversityDepartment of Biotechnology, Kaohsiung Medical UniversityDepartment of Biochemistry, College of Medicine, Kaohsiung Medical UniversityInstitute of Biopharmaceutical Sciences, National Sun Yat-sen UniversityDepartment of Biotechnology, Kaohsiung Medical UniversityDepartment of Biomedical Science and Environmental Biology, Kaohsiung Medical UniversityAbstract Background Camptothecin (CPT) derivatives are widely used in cancer therapies, but their efficacy can be attenuated by resistance mechanisms such as autophagy. We recently showed that the aniline compound 4-[4-(4-aminophenoxy)-2,3,5,6-tetrafluorophenoxy] aniline (TFPA) can potently increase CPT cytotoxicity against non-small cell lung cancer (NSCLC) cells. The purpose of this study was to evaluate whether TFPA improves CPT-based chemotherapy by modulating autophagy and other cell death pathways in NSCLC models. Methods Two NSCLC cell lines, A549 and H1299, were tested. The synergism of CPT and TFPA was evaluated by trypan blue exclusion and colony formation assays. Annexin V staining was used for the detection of apoptosis, and autophagy was assessed by acridine orange staining and immunofluorescence. Flow cytometry-based dihydroethidium staining was used to assess oxidative stress. Changes in the expression of apoptosis-associated factors and autophagy-associated factors were determined by Western blot assays. The synergism of CPT and TFPA was validated using a zebrafish xenograft assay. Results The accumulation of markers for lysosomal expansion (LAMP2) and degradation (cathepsin D) and markers for autophagosome formation (LC3B-II) suggested that blockage of autolysosome formation might impair autophagy in CPT-treated NSCLC cells and subsequently lead to autophagic cell death. Cotreatment with TFPA and CPT induced cell death by increasing the production of reactive oxygen species, which contributed to autophagic impairment and eventually apoptotic cell death in NSCLC cells. Conclusions Our present work suggests that increased autophagic impairment induced by the combination of CPT and TFPA contributes to the apoptotic cell death of lung cancer cells.https://doi.org/10.1186/s12935-025-03657-6Lung cancerAutophagy impairmentCPTAnilineTFPAOxidative stress
spellingShingle Han-Lin Chou
I-Ling Lin
Yei-Tsung Chen
Wen-Tsan Chang
Ann Yu
Wei-Chun Chen
Chang-Yi Wu
Shean-Jaw Chiou
Chih-Wen Shu
Chien-Chih Chiu
Pei-Feng Liu
Aniline TFPA enhances camptothecin-induced anti-NSCLC by modulating oxidative stress and impairing autophagy
Cancer Cell International
Lung cancer
Autophagy impairment
CPT
Aniline
TFPA
Oxidative stress
title Aniline TFPA enhances camptothecin-induced anti-NSCLC by modulating oxidative stress and impairing autophagy
title_full Aniline TFPA enhances camptothecin-induced anti-NSCLC by modulating oxidative stress and impairing autophagy
title_fullStr Aniline TFPA enhances camptothecin-induced anti-NSCLC by modulating oxidative stress and impairing autophagy
title_full_unstemmed Aniline TFPA enhances camptothecin-induced anti-NSCLC by modulating oxidative stress and impairing autophagy
title_short Aniline TFPA enhances camptothecin-induced anti-NSCLC by modulating oxidative stress and impairing autophagy
title_sort aniline tfpa enhances camptothecin induced anti nsclc by modulating oxidative stress and impairing autophagy
topic Lung cancer
Autophagy impairment
CPT
Aniline
TFPA
Oxidative stress
url https://doi.org/10.1186/s12935-025-03657-6
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