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Total proteomic analysis of peripheral blood mononuclear cells from oral squamous cell carcinoma patients using DIA-MS approach

ABSTRACT: The present study explores DIA-MS method to identify potential protein targets in peripheral blood mononuclear cells (PBMC) of oral squamous cell carcinoma patients in a cancer-control cohort. The total proteomic analysis has identified about 3139 proteins among which 449 proteins showed altered expression in OSCC group compared to the control. Within the subset of the differentially expressed proteins, several potential protein candidates were identified with immunomodulatory activities that are favorable towards the cancer progession and survival. Further researach can be carried out to validate and utilize these protein targets in the clinical settings.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood

DISEASE(S): Oral Squamous Cell Carcinoma

SUBMITTER: Keshava Prasad T. S.

LAB HEAD: Dr. T. S. Keshava Prasad

PROVIDER: PXD051789 | Pride | 2024-08-22

REPOSITORIES: Pride

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Project description:A role of vitamin C (ascorbic acid) as an antioxidant molecule has been recognized, largely based on in vitro studies. However, more recently, the concept of M-bM-^@M-^\antioxidant moleculeM-bM-^@M-^] has been reconsidered and its biological function is no longer considered to be simply due to its ability to act as M-bM-^@M-^\electron donorsM-bM-^@M-^], rather, it appears to act by modulating signaling and gene expression. In order to gain a better understanding of the effects of vitamin C supplementation on gene expression in both physiological and pro-inflammatory conditions, we performed a pilot explorative intervention study based on the utilization of microarray and qPCR technologies, designed on a M-bM-^@M-^\in vivo-ex vivoM-bM-^@M-^] structure Five healthy subjects (three males and 2 females, aged 25-40) were recruited for a short term (five days) intervention study. Subjects were asked to assume one tablet of RedoxonM-BM-. (Bayer), a day (containing 1 gr of ascorbate), for 5 days. A baseline blood withdrawal was performed in fasting condition, just before the first vitamin C tablet. Tablets were provided to the volunteers, every morning at the same hour for the following 4 days of the study. The last tablet was supplemented about 1,5 hour before the second and last blood withdrawal, performed again in fasting condition. Plasma and peripheral blood mononuclear cells (PBMNC) were obtained . PBMNC from each subject were splitted in three aliquots: one was stored in lysis buffer at -80M-BM-0C and utilized to isolate RNA for the Affymetrix gene array analysis. The other two aliquots were resuspended in RPMI 1640 w/o phenol red containing 10% of autologous plasma: one was incubated with 10 ug/ml lipopolysaccharides (LPS) for 5 hours at 37M-BM-0C. At the end of the incubation time, the medium was collected and stored at -80M-BM-0C for the assessment of cytokine release in response to the inflammatory stimuli. RNA was extracted from the pelleted cells for gene expression analysis, using the Human NFkB Signaling 96 StellARrayM-bM-^DM-" qPCR array (Lonza). The results obtained with PBMNC isolated after vitamin C supplementation (supplemented PBMNC) were compared to those obtained with PBMNC isolated before the supplementation (baseline PBMNC), for each subject.

			Action	DRS
	PBMC_Cancer_DIA_R1.raw	Raw
	PBMC_Cancer_DIA_R2.raw	Raw
	PBMC_Cancer_DIA_R3.raw	Raw
	PBMC_Normal_DIA_R1.raw	Raw
	PBMC_Normal_DIA_R2.raw	Raw

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Total proteomic analysis of peripheral blood mononuclear cells from oral squamous cell carcinoma patients using DIA-MS approach

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