ABSTRACT
Objective: To observe the platinum drugs resistance effect of N-acetyltransferase 10 (NAT10) overexpression in breast cancer cell line and elucidate the underlining mechanisms. Methods: The experiment was divided into wild-type (MCF-7 wild-type cells without any treatment) group, NAT10 overexpression group (H-NAT10 plasmid transfected into MCF-7 cells) and NAT10 knockdown group (SH-NAT10 plasmid transfected into MCF-7 cells). The invasion was detected by Transwell array, the interaction between NAT10 and PARP1 was detected by co-immunoprecipitation. The impact of NAT10 overexpression or knockdown on the acetylation level of PARP1 and its half-life was also determined. Immunostaining and IP array were used to detect the recruitment of DNA damage repair protein by acetylated PARP1. Flow cytometry was used to detect the cell apoptosis. Results: Transwell invasion assay showed that the number of cell invasion was 483.00±46.90 in the NAT10 overexpression group, 469.00±40.50 in the NAT10 knockdown group, and 445.00±35.50 in the MCF-7 wild-type cells, and the differences were not statistically significant (P>0.05). In the presence of 10 μmol/L oxaliplatin, the number of cell invasion was 502.00±45.60 in the NAT10 overexpression group and 105.00±20.50 in the NAT10 knockdown group, both statistically significant (P<0.05) compared with 219.00±31.50 in wild-type cells. In the presence of 10 μmol/L oxaliplatin, NAT10 overexpression enhanced the binding of PARP1 to NAT10 compared with wild-type cells, whereas the use of the NAT10 inhibitor Remodelin inhibited the mutual binding of the two. Overexpression of NAT10 induced PARP1 acetylation followed by increased PARP1 binding to XRCC1, and knockdown of NAT10 expression reduced PARP1 binding to XRCC1. Overexpression of NAT10 enhanced PARP1 binding to LIG3, while knockdown of NAT10 expression decreased PARP1 binding to LIG3. In 10 μmol/L oxaliplatin-treated cells, the γH2AX expression level was 0.38±0.02 in NAT10 overexpressing cells and 1.36±0.15 in NAT10 knockdown cells, both statistically significant (P<0.05) compared with 1.00±0.00 in wild-type cells. In 10 μmol/L oxaliplatin treated cells, the apoptosis rate was (6.54±0.68)% in the NAT10 overexpression group and (12.98±2.54)% in the NAT10 knockdown group, both of which were statistically significant (P<0.05) compared with (9.67±0.37)% in wild-type cells. Conclusion: NAT10 overexpression enhances the binding of NAT10 to PARP1 and promotes the acetylation of PARP1, which in turn prolongs the half-life of PARP1, thus enhancing PARP1 recruitment of DNA damage repair related proteins to the damage sites, promoting DNA damage repair and ultimately the survival of breast cancer cells.
Subject(s)
Female , Humans , Breast Neoplasms/enzymology , Cell Line, Tumor , Drug Resistance, Neoplasm , MCF-7 Cells , N-Terminal Acetyltransferases/metabolism , Organoplatinum Compounds/pharmacology , Oxaliplatin/pharmacology , X-ray Repair Cross Complementing Protein 1ABSTRACT
Objective To evaluate the effectiveness of psychological health intervention in the elderly urban residents living on minimum subsistence allowances, and to search for a suitable psychological health intervention strategy. Methods The 112 elderly persons living on minimum subsistence allowances in Xuzhou were selected by stratified cluster sampling, and they were matched with 112 controls. The elderly persons living on minimum subsistence allowances were divided into intervention group and control group. A general intervention based on community involved psychological health and care lecture, psychodrama treatment and psychological consultancy was made on the intervention. The SCL-90 was used to evaluate the psychological health status and effectiveness of psychological health intervention. Results The elderly persons living on minimum subsistence allowances had higher SCL-90 scores compared with other residents, and their psychological health status was serious. After the psychological health intervention, the SCL-90 scores were all lower in intervention group than in control group [ interpersonal relationship sensitiveness: (1.13 ± 0. 39) vs.(1.26±0.26), t=2.12, P=0.04; gloom: (1.23±0.66) vs. (1.43±0.24), t=2.08, P=0.04;anxiety: (1. 18±0.50) vs. (1.38±0.34), t=2.17, P=0.03; hostility: (1.24±0.49) vs. (1.40±0.28), t=2.03, P=0.04; other factor: (1. 31±0.56) vs. (1.49±0.31), t=2.04, P=0.04; total score: (115.89± 17.21) vs. (122.64± 10.41), t=2.42, P=0.02]. Conclusions The psychological health parameters are improved by psychological health intervention.