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. 2018 Nov 29;18(1):314.
doi: 10.1186/s12906-018-2383-6.

Proteomic Profiling of Senescent Human Diploid Fibroblasts Treated With Gamma-Tocotrienol

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Free PMC article

Proteomic Profiling of Senescent Human Diploid Fibroblasts Treated With Gamma-Tocotrienol

Jen-Kit Tan et al. BMC Complement Altern Med. .
Free PMC article

Abstract

Background: Replicative senescence of human diploid fibroblasts (HDFs) has been used as a model to study mechanisms of cellular aging. Gamma-tocotrienol (γT3) is one of the members of vitamin E family which has been shown to increase proliferation of senescent HDFs. However, the modulation of protein expressions by γT3 in senescent HDFs remains to be elucidated. Therefore, this study aimed to determine the differentially expressed proteins (DEPs) in young and senescent HDFs; and in vehicle- and γT3-treated senescent HDFs using label-free quantitative proteomics.

Methods: Whole proteins were extracted and digested in-gel with trypsin. Peptides were detected by Orbitrap liquid chromatography mass spectrometry. Mass spectra were identified and quantitated by MaxQuant software. The data were further filtered and analyzed statistically using Perseus software to identify DEPs. Functional annotations of DEPs were performed using Panther Classification System.

Results: A total of 1217 proteins were identified in young and senescent cells, while 1218 proteins in vehicle- and γT3-treated senescent cells. 11 DEPs were found in young and senescent cells which included downregulation of platelet-derived growth factor (PDGF) receptor beta and upregulation of tubulin beta-2A chain protein expressions in senescent cells. 51 DEPs were identified in vehicle- and γT3-treated senescent cells which included upregulation of 70 kDa heat shock protein, triosephosphate isomerase and malate dehydrogenase protein expressions in γT3-treated senescent cells.

Conclusions: PDGF signaling and cytoskeletal structure may be dysregulated in senescent HDFs. The pro-proliferative effect of γT3 on senescent HDFs may be mediated through the stimulation of cellular response to stress and carbohydrate metabolism. The expressions and roles of these proteins in relation to cellular senescence are worth further investigations. Data are available via ProteomeXchange with identifier PXD009933.

Keywords: Gamma-tocotrienol; Human diploid fibroblasts; Proteomics; Replicative senescence.

Conflict of interest statement

Ethics approval and consent to participate

Ethical approval was obtained from UKM Ethical Committee with the approval code: FF-249-2012. Sample collection was conducted according to the UKM Guidelines for Ethical Review of Clinical Research or Research Involving Human subjects 2014. Written informed consent was provided to the participants’ parents.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Graphical representations of differentially expressed proteins (DEPs). Volcano plot of total proteins for (a) young versus senescent cells and (b) vehicle- versus γT3-treated senescent cells. DEPs were highlighted in red. Heat map of DEPs for (c) young versus senescent cells and (d) vehicle- versus γT3-treated senescent cells. Y1–3: young cells; S1–3: senescent cells U1–3: vehicle-treated senescent cells; G1–3: γT3-treated senescent cells; N = 3 biological replicates for each group. Scale bar: young cells were set as control group for senescent cells; vehicle-treated senescent cells were set as control group for γT3-treated senescent cells
Fig. 2
Fig. 2
Distribution of pathways based on Panther Classification System. Pathways related to differentially expressed proteins in (a) young versus senescent cells and (b) vehicle- versus γT3-treated senescent cells. The number in bracket indicates number of proteins

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