Benzofuran-isatin derivative 5d acts as a dual-action agent in colorectal cancer cells by targeting EMT and mitochondrial apoptosis pathways

Abstract

Objective: To investigate the anticancer effects of a novel benzofuran-isatin conjugate [N’-(5-methoxy-2-oxoindolin-3-ylidene)-3-methylbenzofuran-2-carbohydrazide (conjugate 5d)] against human colorectal adenocarcinoma (CRC) HT29 and metastatic SW620 colorectal cancer cells.

Methods: The cytotoxic properties of conjugate 5d were evaluated using the MTT assay. Its anti-oncogenic effects were assessed by real-time monitoring of cell proliferation, migration, and invasion, and by performing a clonogenic assay. Flow cytometry was also used to assess the apoptotic status and cell cycle. Apoptosis, cell cycle, and epithelial-mesenchymal transition-related protein and gene expression levels were also measured.

Results: Conjugate 5d exhibited cytotoxic effects on both CRC cells and enhanced the cytotoxic efficacy of 5-fluorouracil, irinotecan, and oxaliplatin. Conjugate 5d also induced apoptosis by modulation of anti-apoptotic (i.e., Bcl-xl) and pro-apoptotic (i.e., Bax, p53, cytochrome c) proteins, and MMP loss. Docking studies predicted molecular interactions of conjugate 5d with anti-apoptotic Bcl-2, revealing conjugate 5d as a potential Bcl-2 inhibitor. Regarding the oncogenic process, conjugate 5d inhibited CRC cell proliferation, migration, invasion, and colony formation, upregulated E-cadherin expression, and downregulated N-cadherin expression.

Conclusions: Conjugate 5d shows significant anticancer effects against HT29 and SW620 cells by exerting pro-apoptotic and antimetastatic activities. It also enhances the cytotoxic efficacy of conventional chemotherapeutic drugs in CRC cell lines. However, in vivo studies should be conducted to further confirm its efficacy.

Keywords

Colorectal cancer / Synthetic benzofuranisatin conjugate / Apoptosis / Cell cycle / Anticancer / Epithelialmesenchymal transition

Conflict of interest statement

The authors declare that there is no potential conflict of interest.

Funding

This work was supported by Ongoing Research Funding Program, (ORF-2026-344), King Saud University, Riyadh, Sausi Arabia.

Data availability statement

The data supporting the findings of this study are available from the corresponding author upon request.

Authors’ contributions

Conceptualization or design of the work was done by MHA, MAVM and SMN. MM, NSA, TT, WME, AAO, and KAK performed data collection. WME, MAVM and MHA contributed to the methodology. RS contributed to study design, methodology and writing the manuscript. MHA, MAVM, SMN and KAK were involved in data analysis and interpretation. MHA, MAVM and SMN were responsbile for writing and editing the article. All authors have read and approved the final version of the article.

Publisher’s note

The Publisher of the Journal remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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