Downregulation of PER2 Promotes Tumor Progression by Enhancing Glycolysis via the Phosphatidylinositol 3-Kinase/Protein Kinase B Pathway in Oral Squamous Cell Carcinoma.

PURPOSE

PER2 gene expression is downregulated in oral squamous cell carcinoma (OSCC) and may have a pivotal role in tumor suppression. However, the biological function and mechanism of action of PER2 in OSCC remain unclear. In this study, the biological functions and anticancer mechanisms of PER2 in OSCC were investigated.

MATERIALS AND METHODS

Both stably overexpressed and silenced PER2 OSCC cells were established as an experimental group; empty vector lentivirus and scramble short hairpin RNA lentivirus transfected-cells, as negative control groups; and untreated OSCC cells, as a blank group. Cell proliferation, apoptosis, and glycolysis potential assays were conducted. In addition, the expression of phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), phosphorylation of protein kinase B, hexokinase 2 (HK2), pyruvate kinase M (PKM2), and lactate dehydrogenase A (LDHA) was quantified by real-time quantitative polymerase chain reaction and Western blotting. Rescue experiments were performed by the addition of AKT activators in the overexpressed cell line and by the addition of glycolysis inhibitor in the silenced cell line. These findings were verified in vivo using stably transfected OSCC cells overexpressing PER2 implanted in nude mice.

RESULTS

PER2 overexpression significantly inhibited OSCC cell proliferation and glycolysis, promoted cell apoptosis, and reduced the expression of PI3K, phosphorylation of protein kinase B, HK2, PKM2, and LDHA. The converse was observed in PER2-silenced OSCC cells. After the addition of AKT activator to cultures of PER2-overexpressed OSCC cells, reduced glucose uptake, lactic acid production, and cell proliferation, combined with increased apoptosis, were substantially reversed. In addition, after the addition of HK2 inhibitor to PER2-silenced OSCC cells to inhibit glycolysis, the reduction in apoptosis and increased proliferation were significantly countermanded. Tumorigenesis experiments in vivo also confirmed that PER2 overexpression suppressed OSCC growth and decreased the expression of HK2, PKM2, and LDHA.

CONCLUSIONS

PER2 heightened glycolysis via the PI3K/AKT pathway, heightened cell proliferation and inhibited apoptosis via glycolysis, thereby promoting the development of OSCC.