Differential transmission of the molecular signature of RBSP3, LIMD1 and CDC25A in basal/ parabasal versus spinous of normal epithelium during head and neck tumorigenesis: A mechanistic study

Abstract

Head and neck squamous cell carcinoma (HNSCC) is a global disease and mortality burden, necessitating the elucidation of its molecular progression for effective disease management. The study aims to understand the molecular profile of three candidate cell cycle regulatory genes, RBSP3, LIMD1 and CDC25A in the basal/ parabasal versus spinous layer of normal oral epithelium and during head and neck tumorigenesis. Immunohistochemical expression and promoter methylation was used to determine the molecular signature in normal oral epithelium. The mechanism of alteration transmission of this profile during tumorigenesis was then explored through additional deletion and mutation in HPV/ tobacco etiological groups, followed byclinico-pathological correlation. In basal/parabasal layer, the molecular signature of the genes was low protein expression/ high promoter methylation of RBSP3, high expression/ low methylation of LIMD1 and high expression of CDC25A. Dysplastic epithelium maintained the signature of RBSP3 through high methylation/ additional deletion with loss of the signatures of LIMD1 and CDC25A via deletion/ additional methylation. Similarly, maintenance and / or loss of signature in invasive tumors was by recurrent deletion/ methylation. Thus, differential patterns of alteration of the genes might be pre-requisite for the development of dysplastic and invasive lesions. Etiological factors played a key role in promoting genetic alterations and determining prognosis. Tobacco negative HNSCC patients had significantly lower alterations of LIMD1 and CDC25A, along with better survival among tobacco negative/ HPV positive patients. Our data suggests the necessity for perturbation of normal molecular profile of RBSP3, LIMD1 and CDC25A in conjunction with etiological factors for head and neck tumorigenesis, implying their diagnostic and prognostic significance.

Head and Neck Cancers Are Not Alike When Tarred with the Same Brush: An Epigenetic Perspective from the Cancerization Field to Prognosis

“…In opposition to the hypomethylation profile observed in the EWAS, CYP1A1 hypermethylation was more frequently observed in HNSCC from smokers relative to healthy tissues from controls [63]. When promoter methylation and gene deletion of RBSP3, LIMD1, and CDC25A were analyzed together in HNSCC, the frequency of alterations was higher in smokers than in nonsmokers [64].…”

Serum GDF15, a Promising Biomarker in Obese Patients Undergoing Heart Surgery

“…Representative atrial appendage samples ( n = 2 for each group of normal, pre-obese, obese class I, class II, and class III patients) were used for GDF15 protein expression and localization. Paraffin sections (5 μm) were deparaffinized, serially hydrated and stained with Hematoxylin and Eosin for histology ( 14 ). Sections from the same tissue were then used for immunostaining of GDF15.…”

“…Sections from the same tissue were then used for immunostaining of GDF15. In brief, paraffin sections were hydrated, antigens retrieved using Citrate buffer, blocked with 15% goat serum and incubated with anti-GDF15 primary antibody at 1:100 dilution (Santacruz Biotechnology) (14). After incubation with Horse Radish Peroxidase (HRP)-conjugated secondary antibody (1:500), slides were developed with 3,3 ′ Diaminobenzidine (DAB) (Sigma Millipore, MA, US) and counter-stained with Hematoxylin.…”