Elp3 Can Reduce The Extent Of DNA Damage Casused By⁶⁰Coγ In Hepatocellular Carcinoma Cells

Objective:To explore the relationship between Elp3 and DNA damage repair induced by 60Co γ ray and the molecular mechanism of Elp3 with respect to DNA damage repair in hepatocellular carcinoma cells and normal hepatic cells.Methods:1.Contruction of lentiviral vector carrying Elp3 gene.2.Using lentiviral to contract Hep3B/7701 cell lines which overexpression/knock-down of Elp3 stably.3.After the irradiation of 60Co γ ray,the change of DNA damage in different groups was detected by comet assay with OTM as an indicator.4.Cell apoptosis rate in each cell line was measured by flow cytometry.5.Real-time PCR was used to test the expression of the gene related to DNA damage and repair.6.Western blot was used to determine the protein expression with regard to DNA damage and repair.Results:1.We confirmed the lentiviral vector carrying Elp3 gene was contructed successfully after determined by PCR and DNA sequencing.2.We confirmed Hep3B/7701 cell lines which overexpress/knockdown of Elp3 stably was contructed successfully after determined by Real-Time PCR and Western Blot.3.We found Elp3 can reduce the extent of DNA damage in Hep3B but has no effort on 7701 after determined by comet assay.4.After the irradiation of 60Co γ ray,Flow cytometry showed that Elp3 can lower cell apoptosis rate in Hep3B,but Elp3 has no effort on 7701.5.Real-time PCR showed that Elp3 can promote the transcription of DNA-PKcs and Mre11 in Hep3B,but has no effort on the transcription of ATM in Hep3B.Elp3 has no effort on the transcription of DNA-PKcs,Mre11 and ATM in 7701.6.Western Blot showed that Elp3 can promote the protein expression of DNA-PKcs and Mrell in Hqp3B,Elp3 can promote the phosphorylation of AKT and ATM in Hep3B.But Elp3 has no effort on the protein of DNA-PKcs/Mrell/p-AKT/p-ATM in 7701.Conclusion:1.After the irradiation of 60Coγ ray,Elp3 can reduce the extent of DNA damage and can decrease cell apoptosis rate in Hep3B.2.Elp3 reduced the extent of DNA damage in Hep3B may through some factors related to DNA damage and repair,such as DNA-PKcs,AKT,Mrel 1,ATM.3.After the irradiation of 60Co γ ray,Elp3 has no effort on the extent of DNA damage and cell apoptosis rate in 7701,Elp3 has no effort on the expression of DNA-PKcs,p-AKT,Mrel l,p-ATM in 7701.