Investigation of circularRNA Molecules as Biomarkers in Head and Neck Squamous Cell Carcinoma Cell Culture
Project no. 22423 · Status: Kapanmış (Yayın bekleniyor) · 2021
Summary
<p>Head and neck squamous cell carcinomas (HNSCC) are the sixth most common cancer worldwide, affecting approximately 600,000 individuals annually and resulting in over 300,000 deaths. These carcinomas primarily affect the oral cavity, oropharynx, larynx, nasopharynx, and hypopharynx regions. Currently, a multi-modal approach involving various combinations of surgery, radiotherapy, and chemotherapy regimens is employed in treatment. Advances in treatment options have slightly improved both short- and long-term survival rates in certain groups of head and neck cancers. However, local recurrences and secondary primary head and neck tumors remain significant challenges for clinicians. There are currently no biomarkers available that can facilitate early diagnosis, alter treatment options, influence the duration and frequency of patient follow-up, or be used for early recurrence or prognosis determination in head and neck cancers.</p><p>Circular RNA (circRNA) molecules, due to their widespread expression profiles and unusually stable structures, can be easily detected in body fluids, making them potential biomarkers. Recently, they have garnered attention as a new research focus in molecular biology. The circRNAs synthesized in human oral squamous carcinoma cell lines are likely to play significant roles in head and neck cancers and hold potential for clinical use as biomarkers.</p><p>This proposed project investigated all circRNAs produced by human oral squamous carcinoma cell cultures and healthy tissues using next-generation sequencing technology. Initially, human oral squamous carcinoma and healthy cells were cultured. Subsequently, radioresistant carcinoma cells were developed. The radioresistant nature of these cells was confirmed through a colony formation test. Total RNA isolation was then performed from the three cell lines. The purity, concentration, and integrity of the isolated RNA were measured. CircRNA enrichment and linearization were subsequently carried out. A cDNA library was then constructed and sequenced using next-generation sequencing technology. All data were evaluated using bioinformatic analyses, and differentially expressed circRNAs (DEcRs) were identified. Molecular pathway analyses were conducted to explore the roles of the identified DEcRs in cancer development and radioresistance mechanisms. Comparative analyses were performed to identify circRNAs that could serve as potential biomarkers.</p><p>Analyses identified 354 DEcR genes in the three experimental groups (Detroit cells, Detroit-RR cells, and WI38). Among these, 80 circRNA genes were found to be common, with most showing significant differential expression. Sixteen circRNA biomarker candidates were identified through the comparative analysis of sequencing data from Detroit and WI38 cells. These circRNAs exhibited significantly reduced expression in HNSCC cells, with one showing a significant increase. This finding indicates the presence of potential biomarker candidates for HNSCC diagnosis. Through comparative analysis of Detroit and Detroit-RR cells, 37 circRNA biomarker candidates were identified. Ten circRNA genes showed significantly reduced expression levels in radioresistant cells, while 27 circRNA genes exhibited significantly increased expression levels. This highlights potential biomarker candidates that could indicate treatment methods or radioresistant/sensitive conditions before or during the treatment of HNSCC.</p><p>Molecular signaling pathway analyses revealed an enrichment in receptor binding in Detroit cells. In radioresistant Detroit cells, enrichment was observed in the ERBB3 signaling mechanism and receptor binding capacity, enzyme binding, nucleoplasm, nuclear lumen, and microtubule cytoskeleton-related molecular pathways.</p><p>In conclusion, these results demonstrate the presence of potential biomarker candidates that could be utilized in the diagnosis and treatment management of HNSCC patients. Accurate diagnosis of HNSCC patients is crucial, alongside the importance of their treatment and follow-up. Identifying biomarkers that can indicate treatment methods or radioresistant/sensitive conditions before or during treatment is a significant need in this context.</p>