Abstract
Cancer stem cells (CSCs) have garnered attention with their potential for early diagnosis and prognosis of oral squamous cell carcinoma (OSCC). It is still indistinct whether CSCs are recognized with a specific set of characteristics. The present study aimed to assess the association of CD44 with stemness-related, Epithelial Mesenchymal Transition EMT-related genes and the secretome of the CSCs. The single-cell suspension from primary OSCC tumors was prepared by enzymatic digestion and the cells were cultured in-vitro. The cancer stem cells were isolated by CD44+ selection using magnetic cell-sorting. The expression of CD44, proliferation rate, gene expression of EMT-related transcription factors, stemness markers, cytokine levels and angiogenic factors in both cell population was assessed. The sorted CD44+ cells showed significantly higher proliferation rate than heterogenous population. The CD44 expression was >90% in the sorted cells which was higher than the heterogenous cells. The CD44+ CSCs cells demonstrated significant increased levels of EMT-related genes TWIST1 and CDH2 (N-cadherin), CSC-related genes CD44 and CD133 (PROM1), stemness-related genes OCT4, SOX2, inflammatory cytokines IL-1ß, IL-12, IL-18 and TNF-α and angiogenic factors Angiopoietin-1, Angiopoietin-2, bFGF and VEGF while levels of epithelial gene CDH1 (E-cadherin) decreased in comparison to mixed cell population. The genetic and secretome profiling of the CD44+ CSCs could serve as diagnostic and prognostic tools in the treatment of oral cancers.
Highlights
Head and neck cancers (HNCs) are predominant in Asia and oral squamous cell carcinoma (OSCC) in particular accounts for nearly 90 percent of cancer-related fatalities [1]
The severity of dysplastic changes in premalignant lesions are concomitant with inferior clinical consequences, but more demanding approaches are required to envisage the tumor forming prospects of these premalignant lesions with respect to Cancer stem cells (CSCs) and stemness markers
After incubating at 37 ◦C for 30 min, the enzyme action was halted by adding fetal bovine serum (FBS) (Gibco, Rockville, MD, USA); following this, the mixture was passed through a 70-micron cell-strainer (Corning, NY, USA)
Summary
Head and neck cancers (HNCs) are predominant in Asia and oral squamous cell carcinoma (OSCC) in particular accounts for nearly 90 percent of cancer-related fatalities [1]. Diagnosis of cancer-related symptoms is encouraged to understand and develop strategies for treatment of OSCCs. The prime time for medical intervention is the point where cancer stem cell (CSC)-like cells start appearing. Tumors frequently encompass ordered provisions of gene-wide or epigenetic divergent cancerous and non-cancerous cellular processes [5,6]. In these instances, some speculative phenotypes of tumor-forming cells are being identified through exhaustive experimental work with single-cell suspensions from tumors, xenotransplantation studies and phenotypic cell-surface antigens [7,8]. The relationship between proclaimed stem cell markers with the oral epithelial tumorigenicity should be studied thoroughly
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