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Cornulin, a potential prognostic biomarker in head and neck squamous cell carcinoma, acts as an anti-tumour agent by inhibiting cell cycle through upregulation of p18.

ABSTRACT: The xenograft models were developed using 4–6-week-old CD1-Foxn1nu nude male mice and subcutaneously inoculating HNSCC cell lines, i.e., Cal27. In each mouse, the left flank received Cal27 untreated cells and the right flank received Cal27 Cornulin-overexpressing cells to allow within-animal comparison. After tumor development, mice were sacrificed, and xenograft tumors were excised. Tumor tissues were then subjected to protein extraction, tryptic digestion, and LC-MS/MS-based proteomic analysis to identify differentially expressed proteins associated with Cornulin overexpression in vivo.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Epithelial Cell

DISEASE(S): Head And Neck Squamous Cell Carcinoma

SUBMITTER: Arnab Pal

LAB HEAD: Arnab Pal

PROVIDER: PXD065385 | Pride | 2025-07-16

REPOSITORIES: Pride

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Project description:We describe a simple method, Digital Restriction Enzyme Analysis of Methylation (DREAM), based on sequential DNA digestion with a pair of methylation-blocked and methylation-tolerant neoschizomeric restriction enzymes SmaI/XmaI followed by end repair and ultra-deep sequencing. DREAM provides information on 160,000 unique CpG sites of which 39,000 are in CpG islands, and 33,000 are at transcription start sites (-1 kb to +1 kb) of 13,139 RefSeq genes. We compared DNA methylation values in white blood cells from 4 healthy individuals and found them to be remarkably uniform. Interindividual differences >30% were observed only at 227 of 28,331 (0.8%) of autosomal CCCGGG sites covered by 100+ sequencing reads. Similarly, differences at only 59 sites were observed between the cord and adult blood. Conserved methylation patterns in healthy blood cells contrasted with extensive changes affecting 18-40% of CpG sites in leukemia. The method is cost effective, quantitative (r2=0.93 when compared to bisulfite pyrosequencing), reproducible (r2=0.997), and can detect differences >25% with false positive rate <0.001. Accurate analysis of changes in DNA methylation will be useful in quantifying epigenetic effects of environment and nutrition, correlating developmental epigenetic variation with phenotypes, understanding epigenetics of cancer and chronic diseases, measuring the effects of drugs on DNA methylation or deriving new biological insights into mammalian genomes. Digital restriction enzyme analysis of methylation (DREAM) was performed to determine the DNA methylation profiles of healthy white blood cells from cord blood and adult blood, acute myeloid leukemia bone marrow, and two leukemia cell lines (HEL and K562). In this approach, genomic DNA is sequentially cut at CCCGGG sites with the methylation-sensitive enzyme SmaI (blunt ends) and its methylation-tolerant neoschizomer XmaI (5'CCGG overhangs), creating different end sequences that represent methylation status of the CCCGGG sites. These end sequences are analyzed by next-generation sequencing, and thereafter the methylation status at individual CCCGGG sites across the genome can be determined.

			Action	DRS
	1T.msf	Msf
	1T.raw	Raw
	1U.msf	Msf
	1U.raw	Raw
	2T.msf	Msf

Dataset Information

Cornulin, a potential prognostic biomarker in head and neck squamous cell carcinoma, acts as an anti-tumour agent by inhibiting cell cycle through upregulation of p18.

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