Abstract
Introduction
Glioblastoma (GBM), a grade IV intracranial tumour that is widely known for its highly aggressive and heterogeneous nature, contributing to treatment resistance. Tocotrienol-rich fraction (TRF), a natural extract of vitamin E derived from palm oil, has been reported to exhibit potent anti-cancer effects. However, scanty is known on the effect of TRF on GBM, while the underlying molecular mechanism remains unclear. This study aimed to uncover the anti-cancer mechanism exerted by TRF on a resistant human cell model of GBM (LN-18 cell line) at the proteome level.
Material and Methods
The half maximal inhibitory concentration (IC50) of TRF on LN-18 cells was identified from 24 - 72 hrs using a range of concentrations (0-40 µg/mL) treated. Protein was extracted from LN-18 cells following 72 hrs exposure to TRF, then label-free quantitative proteomic profiling was utilized to analyse the proteomic changes and determine the differentially expressed proteins (DEPs) between the TRF-treated and untreated control. The identified DEPs were further analysed using bioinformatic databases to recognise functional classification of proteins and pathway enrichment analysis.
Results and Discussions
TRF treatment in GBM demonstrated a concentration and time-dependent reduction in cell viability with IC50 values 25, 18 and 15μg/mL for 24, 48 and 72 hrs, respectively. A total of 65 DEPs were identified from the proteome of TRF-treated LN-18 cells as compared to the untreated control. Bioinformatic analysis reveals that 65 DEPs modulated by TRF are involved in the cellular response to stress (CRS) pathways. PANTHER functional analysis demonstrated these DEPs are characterised into metabolic conversion enzymes (11 proteins, mainly in glycolysis process), cytoskeletal proteins (8 proteins), chaperones (4 proteins), chromatin-binding proteins (11 proteins), and translational proteins (7 proteins). Further analysis demonstrated that chaperone (HSPA5), chromatin-binding proteins (H1 and H2B histone proteins) and translational proteins (60S and 40S ribosomal proteins) are found to be associated with the induction of CRS by TRF in LN-18 cells.
Conclusion
The current findings suggest that TRF treatment induced anti-proliferative effect through the modulation of DEPs that are associated with CRS pathways, This is further supported by the alignment of the DEPs to functional classes as chaperones, chromatin-binding proteins and translational proteins which are incorporated to the CRS pathways.
Glioblastoma (GBM), a grade IV intracranial tumour that is widely known for its highly aggressive and heterogeneous nature, contributing to treatment resistance. Tocotrienol-rich fraction (TRF), a natural extract of vitamin E derived from palm oil, has been reported to exhibit potent anti-cancer effects. However, scanty is known on the effect of TRF on GBM, while the underlying molecular mechanism remains unclear. This study aimed to uncover the anti-cancer mechanism exerted by TRF on a resistant human cell model of GBM (LN-18 cell line) at the proteome level.
Material and Methods
The half maximal inhibitory concentration (IC50) of TRF on LN-18 cells was identified from 24 - 72 hrs using a range of concentrations (0-40 µg/mL) treated. Protein was extracted from LN-18 cells following 72 hrs exposure to TRF, then label-free quantitative proteomic profiling was utilized to analyse the proteomic changes and determine the differentially expressed proteins (DEPs) between the TRF-treated and untreated control. The identified DEPs were further analysed using bioinformatic databases to recognise functional classification of proteins and pathway enrichment analysis.
Results and Discussions
TRF treatment in GBM demonstrated a concentration and time-dependent reduction in cell viability with IC50 values 25, 18 and 15μg/mL for 24, 48 and 72 hrs, respectively. A total of 65 DEPs were identified from the proteome of TRF-treated LN-18 cells as compared to the untreated control. Bioinformatic analysis reveals that 65 DEPs modulated by TRF are involved in the cellular response to stress (CRS) pathways. PANTHER functional analysis demonstrated these DEPs are characterised into metabolic conversion enzymes (11 proteins, mainly in glycolysis process), cytoskeletal proteins (8 proteins), chaperones (4 proteins), chromatin-binding proteins (11 proteins), and translational proteins (7 proteins). Further analysis demonstrated that chaperone (HSPA5), chromatin-binding proteins (H1 and H2B histone proteins) and translational proteins (60S and 40S ribosomal proteins) are found to be associated with the induction of CRS by TRF in LN-18 cells.
Conclusion
The current findings suggest that TRF treatment induced anti-proliferative effect through the modulation of DEPs that are associated with CRS pathways, This is further supported by the alignment of the DEPs to functional classes as chaperones, chromatin-binding proteins and translational proteins which are incorporated to the CRS pathways.
| Original language | English |
|---|---|
| Pages (from-to) | 141-142 |
| Number of pages | 2 |
| Journal | Molecular Oncology |
| Volume | 17 |
| Issue number | S1 |
| DOIs | |
| Publication status | Published - Jun 2023 |
| Event | Annual Congress of the European Association for Cancer Research (EACR 2023): Innovative Cancer Science - Lingotto Fiere, Torino, Italy Duration: 12 Jun 2023 → 15 Jun 2023 https://febs.onlinelibrary.wiley.com/toc/18780261/2023/17/S1 https://www.eacr2023.org/ |
