Transcriptomic Analysis of the Anticancer Effects of Annatto Tocotrienol, Delta-Tocotrienol and Gamma-Tocotrienol on Chondrosarcoma Cells

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.

Molecular Mechanism of Tocotrienol-Mediated Anticancer Properties: A Systematic Review of the Involvement of Endoplasmic Reticulum Stress and Unfolded Protein Response

“…Additionally, δ-tocotrienol also induced prostate tumor cell paraptosis with extensive cytoplasmic vacuolation [52]. A similar finding was reported, whereby annatto tocotrienol, γand δ-tocotrienols induced paraptosis or paraptosis-like cell death with extensive cytoplasmic vacuolation in osteosarcoma SW1353 cells [31], human melanoma A375 cells [61], and human colon cancer SW620 cells [62,63]. Furthermore, pre-treatment with salubrinal (anERS inhibitor) significantly suppressed δ-tocotrienol-mediated cytoplasmic vacuolation [52], which further suggests the importance of ERS in executing paraptosis.…”

“…Nutrients 2023, 15, x FOR PEER REVIEW 4 of 18 in several cancerous cells originating from the mammary gland, digestive tract, liver, prostate, lung, and bone cancer [24,[30][31][32][33][34][35]. Mechanistically, the anticancer properties of tocotrienol are subjected to the types of isoforms and cancer cell models.…”

“…Tocotrienol is known to inhibit 3-hydroxy-3-methyglutaryl-CoA (HMG CoA) reductase activity and expression to exert cholesterollowering effects [27][28][29]. Additionally, tocotrienol is widely reported to have anti-cancer properties, whereby it induces growth arrest and cell death, primarily apoptosis, in several cancerous cells originating from the mammary gland, digestive tract, liver, prostate, lung, and bone cancer [24,[30][31][32][33][34][35]. Mechanistically, the anticancer properties of tocotrienol are subjected to the types of isoforms and cancer cell models.…”

Role of vitamins A, C, D, E in cancer prevention and therapy: therapeutic potentials and mechanisms of action

“…AnTT, -T3 and -T3 produced G1 arrest in SW1353 cells after 24 h of treatment, which escalated to apoptosis after 48 h. Furthermore, tocotrienol treatment caused considerable cytoplasmic vacuolation in SW1353 cells. The transcriptome analysis after tocotrienol therapy revealed enhanced signaling pathways in the endoplasmic reticulum stress, unfolded protein response, autophagy, and transcription ( 125 ).…”

Therapeutic potential of bixin on inflammation: a mini review

“…Two thirds of the annual production is marketed as dried seeds while the remaining as colorant ( 11 ). Previous studies have documented that the seeds of B. orellana possess various pharmacological properties, including antioxidant ( 13 ), antimicrobial ( 14 ), bone protective ( 15 ), nephroprotective ( 16 ), hypoglycemic ( 17 ), hypocholesterolemia ( 18 ), anticancer ( 19 ), and anti-inflammatory ( 18 ). While the other bioactive compounds from the seeds like tocotrienol have been reviewed extensively, bixin, which is unique to this plant receives less attention for its medicinal benefits.…”