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  • Evaluation of the expression of two circular non-coding transcripts involved in pluripotency and cell proliferation, CircBIRC6 and CircFOXP1, in esophageal cancer cell lines and stem cells.

  • hamideh sayahi,1 Mitra khalili,2,*
    1. Faculty of Medicine, Department of Genetics and Molecular Medicine, Zanjan University of Medical Sciences, Zanjan, Iran
    2. Faculty of Medicine, Department of Genetics and Molecular Medicine, Zanjan University of Medical Sciences, Zanjan, Iran


  • Introduction: Background and Objective Esophageal cancer is one of the rare types of cancer, characterized by a significant increase in mortality rates. This disease is often diagnosed at advanced stages, highlighting the need for new and effective diagnostic and therapeutic methods. In recent years, extensive research has been conducted on non-coding molecules such as circular RNAs (circRNAs), which play important roles in gene expression regulation and biological processes. CircRNAs have been proposed as potential biomarkers for the identification and prediction of cancer, and they can aid in understanding the molecular mechanisms of the disease. The objective of this study is to investigate the expression of circRNAs in esophageal cancer cell lines and compare them with mesenchymal stem cells and embryonal carcinoma. Through this research, we aim to identify specific circRNAs that may serve as new biomarkers for esophageal cancer. This information could lay the groundwork for future research in this area and contribute to the development of novel diagnostic and therapeutic approaches. Overall, this study explores the relationship between circRNAs and esophageal cancer, emphasizing the importance of these molecules in disease detection and prediction.
  • Methods: In this study, several cell lines were selected, including esophageal cancer cell lines, embryonal carcinoma (NT2), mesenchymal stem cells (MSCs), and induced pluripotent stem cells (iPSCs) (NP40-8 and NP41-17), and cultured under appropriate conditions. RNA was extracted from these cell lines at a density of 70 to 80 percent, and the quality and concentration of the RNA were measured using a Nanodrop c2000 device. For cDNA synthesis, 1 microgram of RNA was utilized with the SMOBIO kit. Subsequently, the concentration of primers and annealing temperature for the Real-Time PCR reaction were optimized, and the expression of circular RNAs was investigated in the iPSC cell line. Finally, the expression levels of the RNAs were normalized against a reference gene, and statistical analysis was performed using the Kruskal-Wallis test.
  • Results: In this study, the cell lines were successfully cultured, and their quality was confirmed. The concentration of the extracted RNA ranged from 460 to 2300 ng/µl, with 260/280 and 260/230 ratios indicating acceptable RNA quality. Additionally, cDNA synthesis was successfully achieved, and the expression of circRNAs was confirmed, as the melt curves demonstrated the specificity of the products. Ultimately, circBIRC6 and circFOXP1 showed significant expression in MSC and NT2 cell lines compared to certain esophageal cancer cell lines, confirming their potential as biomarkers for esophageal cancer.
  • Conclusion: This study investigates circular RNAs (circRNAs) in esophageal cancer, focusing on two key circRNAs, circBIRC6 and circFOXP1. This disease is associated with recurrence and resistance to treatment, making the identification of molecular mechanisms for early detection essential. CircBIRC6 enhances the stemness of cancer cells by sequestering miR-34a and miR-145, potentially leading to tumor recurrence. CircFOXP1 supports WNT signaling pathways, and its disruption can contribute to oncogenesis. CircRNAs could serve as new biomarkers for the early detection of esophageal cancer. Further research is necessary to explore the mechanisms of circRNAs and their therapeutic potential. Ultimately, circRNAs offer new opportunities for the diagnosis and treatment of esophageal cancer, aiding in a better understanding of this disease.
  • Keywords: Circular RNAs (circRNAs), esophageal cancer, cell lines, stem cells, Real-Time PCR.

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