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Transcriptomic Analysis of the Anticancer Effects of Annatto Tocotrienol, Delta-Tocotrienol and Gamma-Tocotrienol on
Kok-Lun Pang1, Lian-Chee Foong2, Norzana Abd Ghafar3
1Department of Pharmacology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun Razak, Cheras 56000, Malaysia.
Abstract:
Previous studies have demonstrated the anticancer activities of tocotrienol on several types of cancer, but its effects on chondrosarcoma have never been investigated. Therefore, this study aims to determine the anticancer properties of annatto tocotrienol (AnTT), γ-tocotrienol (γ-T3) and δ-tocotrienol (δ-T3) on human chondrosarcoma SW1353 cells. Firstly, the MTT assay was performed to determine the half-maximal inhibitory concentration (IC50) of tocotrienol on SW1353 cells after 24 h treatment. The mode of cell death, cell cycle analysis and microscopic observation of tocotrienol-treated SW1353 cells were then conducted according to the respective IC50 values. Subsequently, RNAs were isolated from tocotrienol-treated cells and subjected to RNA sequencing and transcriptomic analysis. Differentially expressed genes were identified and then verified with a quantitative PCR. The current study demonstrated that AnTT, γ-T3 and δ-T3 induced G1 arrest on SW1353 cells in the early phase of treatment (24 h) which progressed to apoptosis upon 48 h of treatment. Furthermore, tocotrienol-treated SW1353 cells also demonstrated large cytoplasmic vacuolation. The subsequent transcriptomic analysis revealed upregulated signalling pathways in endoplasmic reticulum stress, unfolded protein response, autophagy and transcription upon tocotrienol treatment. In addition, several cell proliferation and cancer-related pathways, such as Hippo signalling pathway and Wnt signalling pathway were also significantly downregulated upon treatment. In conclusion, AnTT, γ-T3 and δ-T3 possess promising anticancer properties against chondrosarcoma cells and further study is required to confirm their effectiveness as adjuvant therapy for chondrosarcoma.
Insights
Annatto tocotrienol (AnTT), γ-tocotrienol (γ-T3), and δ-tocotrienol (δ-T3) show anticancer effects on chondrosarcoma cells. These compounds induce cell cycle arrest and apoptosis, suggesting potential as chondrosarcoma therapies.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Tocotrienols exhibit anticancer activities across various cancer types.
- The effects of tocotrienols on chondrosarcoma remain unexplored.
- Chondrosarcoma is a primary bone malignancy with limited treatment options.
Purpose of the Study:
- To investigate the anticancer potential of annatto tocotrienol (AnTT), γ-tocotrienol (γ-T3), and δ-tocotrienol (δ-T3) against human chondrosarcoma SW1353 cells.
- To elucidate the mechanisms underlying tocotrienol-induced cytotoxicity in chondrosarcoma.
- To identify molecular pathways affected by tocotrienol treatment in chondrosarcoma cells.
Main Methods:
- Cell viability was assessed using MTT assays to determine IC50 values.
- Cell death, cell cycle progression, and morphological changes were analyzed.
- RNA sequencing and quantitative PCR were employed for transcriptomic and gene expression analysis.
Main Results:
- AnTT, γ-T3, and δ-T3 induced G1 cell cycle arrest at 24 hours, progressing to apoptosis by 48 hours.
- Cytoplasmic vacuolation was observed in treated chondrosarcoma cells.
- Transcriptomic analysis revealed upregulation of endoplasmic reticulum stress, unfolded protein response, and autophagy pathways, alongside downregulation of Hippo and Wnt signaling pathways.
Conclusions:
- Annatto tocotrienol, γ-tocotrienol, and δ-tocotrienol demonstrate significant anticancer properties against human chondrosarcoma cells.
- The findings suggest that tocotrienols modulate key cellular pathways involved in cancer progression.
- Tocotrienols hold promise as potential adjuvant therapies for chondrosarcoma, warranting further investigation.
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