Study Of SIRT1 Single Nucleotide Polymorphisms On The Effect Of Notch1 Acetylation Modification On Invasion And Migration Of Breast Cancer Cells

Objective: To investigate the molecular mechanism by which acetylation levels of Notch1 protein affect the expression and activity of Notch1 protein in breast cancer cells carrying SIRT1 SNPs site,thereby regulating the invasion and migration of breast cancer cells.Methods:(1)SIRT1-promoter-CDS-MDA-MB231 and SIRT1-promoter-CDS-MCF-7 breast cancer cells were used,each cell contained four cell lines,followed by SIRT1-promoter-CDS-WT cell line,SIRT1-rs3758391-CDS-MUT cell line,SIRT1-rs35706870-CDS-MUT cell line and SIRT1-rs2394443-CDS-MUT cell line,each carrying the corresponding SIRT1 promoter region SNP+coding region(CDS region).(2)Co-IP detects the interaction between SIRT1 and Notch1 protein.(3)Specific anti-acetylated lysine antibody and WB detect the level of Notch1 acetylation modification.(4)WB detected the expression of Notch1 and the active molecule N1 IC protein.(5)Transwell experiment to detect cancer cell invasion and migration ability.Results:(1)The acetylation modification level of Notch1 protein decreased in the SIRT1-rs3758391-MUT group(P<0.005),and the acetylation modification level of Notch1 protein in the SIRT1-rs35706870-MUT and SIRT1-rs2394443-MUT groups increased(P< 0.05;P< 0.005).(2)The expression of Notch1 protein(P < 0.05)and N1 IC protein(P < 0.05)in SIRT1-rs378391-MUT group decreased and Notch1 protein expression in SIRT1-rs35706870-MUT and SIRT1-rs2394443-MUT group(P<0.005;P<0.005)and N1 IC protein expression(P<0.05;P<0.05)were erected.(3)The migration capacity(P<0.005)and invasion capacity(P<0.005)of breast cancer cells in the SIRT1-rs3758391-CDS-MUT group were weakened,and the migration capacity(P< 0.05;P<0.05;P<0.05)and invasion ability(P < 0.05;P < 0.05)of SIRT1-rs35706870-CDS-MUT and SIRT1-rs2394443-CDS-MUT cancer cells were enhanced.Conclusions:(1)When the rs3758391 site mutation in the promoter region of SIRT1 gene was mutated,the acetylation level of Notch1 was reduced,the expression and activity of Notch1 protein were downregulated,and the migration and invasion ability of breast cancer cells were weakened,suggesting that rs3758391 was a protective factor for breast cancer.(2)When the rs35706870 locus and rs2394443 locus mutations in the promoter region of SIRT1 gene,the acetylation level of Notch1 increased,the expression and activity of Notch1 protein were upregulated,and the migration and invasion ability of breast cancer cells were enhanced,suggesting that rs35706870 and rs2394443 were risk factors for breast cancer.