METABOLITE PROFILING, CYTOTOXICITY, AND PRO-APOPTOTIC EVALUATION OF CINNAMOMUM MYRIANTHUM MERR. IN HT-29 COLORECTAL CANCER CELLS

RHIAN JAYMAR RAMIL; YUKIE SATO; KYOKO KOBAYASHI; KENROH SASAKI; FRANCISCO M. HERALDE III

doi:10.22159/ajpcr.2025v18i12.56710

Authors

RHIAN JAYMAR RAMIL The Graduate School, Centro Escolar University, Manila, Philippines. and Department of Pharmacy, College of Health Sciences, Mariano Marcos State University, Batac City, Ilocos Norte, Philippines. https://orcid.org/0000-0003-0729-7671
YUKIE SATO Department of Pharmacognosy, Tohoku Medical and Pharmaceutical University, Sendai, Japan.
KYOKO KOBAYASHI Department of Pharmacognosy, Tohoku Medical and Pharmaceutical University, Sendai, Japan.
KENROH SASAKI Department of Pharmacognosy, Tohoku Medical and Pharmaceutical University, Sendai, Japan.
FRANCISCO M. HERALDE III Department of Biochemistry and Molecular Biology, College of Medicine, University of the Philippines-Manila, Manila, Philippines.

DOI:

https://doi.org/10.22159/ajpcr.2025v18i12.56710

Keywords:

Apoptosis, Cinnamomum myrianthum, Colorectal cancer, Cytotoxicity, metabolomics

Abstract

Objective: To evaluate the metabolite profile as well as the cytotoxic and pro-apoptotic activities of Cinnamomum myrianthum on HT-29 colorectal cancer (CRC) cells in vitro.

Methods: The cytotoxic effects of C. myrianthum crude extract (CmCE) and its fractions were assessed against HT-29 CRC cells using the WST-8 assay. The half-maximal inhibitory concentration (IC50) and selectivity index (SI) were determined. The most bioactive fraction was further subjected to untargeted metabolite profiling using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry. Apoptosis induction was analyzed through Annexin V-FITC/PI staining through flow cytometry, while caspase-3/7 activation was measured using the Caspase-Glo® 3/7 luminescence assay.

Results: CmCE exhibited moderate cytotoxic activity against HT29 CRC cells with an IC50 value of 32.7 μg/mL (p<0.001). Among the tested fractions (Cm1A-Cm1E), Cm1C demonstrated the most potent cytotoxic effect, with an IC50 of 25.7 μg/mL (p< 0.001) and a SI of 3.63. Flow cytometry analysis revealed that Cm1C significantly induced late-stage apoptosis in HT29 cells. In addition, Cm1C markedly increased caspase-3/7 activity with 2.7- fold (p<0.001) at higher concentration, indicating activation of the apoptotic pathway. Metabolite profiling of Cm1C identified several polyphenolic compounds, including cinnamtannin B1, rutin, hyperin, and kaempferol glycosides.

Conclusion: These findings indicated that Cm1C, a polyphenol-rich fraction of C. myrianthum, exhibited selective cytotoxicity against HT29 CRC cells and induced apoptosis through caspase-3/7 activation. This fraction shows promise as a potential natural candidate for CRC therapy.

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