Knockdown of sirtuin6 positively regulates acetylation of DNMT1 to inhibit the NOTCH signaling pathway in non-small cell lung cancer cell lines.

BACKGROUND AND AIM: Sirtuin proteins (1-7) are nicotinamide adenine dinucleotide (NAD)-dependent deacetylases and ADP-ribosyl transferases (class III histone deacetylase enzymes (HDAC)) mainly involved in the removal of the acetyl group from histone proteins. SIRT6, one of the sirtuins, plays a major role in cancer progression in many types of cancer conditions. We recently reported that SIRT6 acts as an oncogene in NSCLC; thus, silencing of SIRT6 inhibits cell proliferation and induces apoptosis in NSCLC cell lines. NOTCH signaling has been reported to be involved in cell survival and regulates cell proliferation and differentiation. However, recent studies from different groups have converged on the notion that NOTCH1 may be an important oncogene in NSCLC. The abnormal expression of NOTCH signaling pathway members is a relatively frequent event in patients with NSCLC. SIRT6 and the NOTCH signaling pathway might play a critical role in tumorigenesis since they are highly expressed in NSCLC. This study has been performed to explore the exact mechanism by which SIRT6 inhibits cell proliferation and induces the apoptosis of NSCLC and its correlation with NOTCH signaling.

MAIN METHODS: In vitro experiments with human NSCLC cells have been performed. Immunocytochemistry study was used to analyze the expression of NOTCH1 and DNMT1 in A549 and NCI-H460 cell lines. RT-qPCR, Western Blot, Methylated DNA specific PCR, and Co-Immunoprecipitation were performed to explore the key events in the regulation of NOTCH signaling by silencing SIRT6 in NSCLC.

KEY FINDINGS: The findings of this study suggest that silencing of SIRT6 significantly promotes the acetylation status of DNMT1 and stabilizes it. Consequently, acetylated DNMT1 translocates into the nucleus and methylates the NOTCH1 promoter region, resulting in the hindering of NOTCH1-mediated NOTCH signaling.