The Protective Effect Of RMP In Genome

Objective:By studying the protective roles of RMP in HepG2cells with p53as the control, toexplore the relationship of RMP and p53in protection of the genome,and the molecularmechanisms of RMP in the protective effect.Methods:1. Comet assay was used to detect the change of DNA damage in different groupsafter the irradiation of60Co γ ray.2. Immunofluorescence(IF) was used to confirm the extent of DNA damage indifferent groups with using γ-H2AX as a DNA damage symbol.3. Real-time PCR was used to detect the gene expression of RMP and p53underthe different treated conditions.4. Western blot was used to detect the protein expression of RMP and P53underthe different treated conditions.5. Real-time PCR was used to detect the expression of the gene related to DNAdamage and repair.Results:1. Comet assay results showed that RMP and p53both can reduce the extent ofDNA damage in HepG2cells.2. IF results showed that the number of cells containing γ-H2AX foci did not havesignificant difference in different groups. But the average number of γ-H2AX foci ineach hepatoma cell was different. These results further confirmed the results of RMPand p53both can reduce the extent of DNA damage，and this results consistent with theconclusions of comet assay.3. The gene expression of RMP and p53detected by Real-time PCR showed thatboth genes have no correlation in gene expression level.4. The protein expression of RMP and P53detected by western blot showed that there is no correlation between RMP and P53under protein expression level.5. Real-time PCR showed that the expression of RAD50、GADD45、BRCA1、CDC25was related to RMP in DNA damage.Conclusion:The study confirmed that both RMP and p53can protect the genome in HepG2cells after the irradiation of60Co γ ray.From the mRNA and protein expression level ofRMP and p53under the different treated conditions,we can concluded that RMP andp53have no correlation in protection of the genome.And from the expression of thegenes related to DNA damage and repair,we may concluded that RMP will increase theviability of hepatoma cells by enhancing its DNA repairing ability.