Autophagy Overactivity Regulate Terf1 and Terf2 in Positive and Negative-Telomerase Cancer Cell Lines
Abstract A recent suggestion for cancer therapy is targeting intracellular homeostatic signaling pathways like autophagy providing the balance between metabolism and cell cycling. Our study focused on investigating the relationship between autophagy activation by Beclin1 transfection and assessing Terf1 and Terf2 expression as shelterin proteins. The beclin1-containing plasmid was introduced to the U-2OS and Huh7 cell lines using Lipofectamine. The LC3-II as an intracellular autophagosomal marker was detected in transfected cells by flow cytometry. Also, the cells were treated with 3-methyladenine and metformin as autophagy inhibitors and inducers, respectively. Finally, the expression levels of Terf1 and Terf2 were analyzed by real-time PCR. Fluorescent images and flow cytometry results proved excellent GFP expression in the transfected cells. The results of real-time PCR demonstrated that autophagy induction by Beclin1 was increased Terf1 expression level in U-2OS cells up to 451%, while Huh7 cells suffered from the decreased expression of Terf1. Altogether, Terf2 expression was enhanced significantly in both cell lines after 48h treatment in comparison with 24h treatment. The obtained data provided that Beclin1-based activation of autophagy leads to overexpression of some protective shelterin proteins.