Growth arrest of human lens epithelial cell SRA01/04 mediated by H_2O_2 conbined with epigenetic modification

Objective To explore the epigenetic modification change under the growth arrest of human lens epithelial cell SRA01/04 mediated by H2O2.Methods The cultured human lens epithelial cells SRA01/04 were divided into 3 groups:Group A or pre-H2O2 group,that is after 0.5 μmol·L-1 H2O2 was added to the cultured SRA01/04 cells for 30 minutes,the cells were co-cultivated with 300 μL Pirenoxine solution for 23.5 hours;Group B or post-H2O2 group,that is after co-culture of the SRA01/04 cells with 300 μL Pirenoxine solution for 23.5 hours,0.5 μmol·L-1 H2O2 was added and continued to cultivate for 30 minute;Group℃or control group,that is SRA01/04 cells were cultured for 24 hours with neither H2O2 nor drug.Each group was detected by immunocytochemical reaction and Western bolt and the expression of histone deacetylase(HDAC)1,nuclear factor-Kappa B(NF-κB),c-myc and Cyclin D1 was analyzed.Results The immunocytochemical reaction showed that the scanned gray-scaled means of NF-κB in group A(192.25±5.56)group B(170.40±2.50)group C(157.14±1.09).The three groups all had statistical(all P0.01);Those of HDAC1,c-myc and CyclinD1 were all group Cgroup Bgroup A and these groups all had statistical significance(all P0.01).The Western bolt showed that compared with control group,HDAC1 activity decreased after treated with H2O2 and group A appeared more clearly than group B;The expression of NF-κB became stronger,which appeared more clearly in group A than that in group B.Conclusion Under the growth arrest of human lens epithelial cell SRA01/04 mediated by H2O2,the HDAC1 activity decreases,NF-κB factor is up-regulated and the expressions of c-myc and CyclinD1 are down ragulated.