The Effect Of SiRNA-mediated Silencing Of ADAR1 Gene On Cell Cycle And Apoptosis In Mixed Mouse Lymphotcyte Culture

RNA editing is a process to remodify the mRNA coding the genes and the enzyme which has the modifying function is called RNA editing enzyme. After transcribing DNA into RNA , RNA editing enzyme changes the gene sequences and the genetic information by deleting, adding or editing the nucleotide in pre-RNA, regulating the receptor-mediated signal transduction and then produces extensive physiological effect.RNA editase transforms or modifies the nucleotide of pre-RNA by deaminization, and hydrolytic deaminize C6 amino group of the adenosine at the specific RNA site, changing adenosine to inosine or changing cytosine to uracil,but inosine was recognized as guanine in mRNA. These substitutions transform the original heredity codon or shearing site, and change the structure and function of the protein they coded. In previous work we have observed that lymphocyte proliferation was inhibited, when the expression of ADAR1 was lowed during rejection.But the role of ADAR1 plays in rejection and the ways how ADAR1 involves in rejection are still unknown.In the present study we describe experiments aimed at the changes of cel1 cycle and apoptosis when ADAR1 expression was degraded in mixed lymphocyte culture.AIM: To investigate the effect of ADAR1 suppressed on cel1 cycle and apoptosis in mixed lymphocyte culture test.METHODS: After specific siRNAs of ADAR1 were transfected into mouse lymphocyte by electroporation for 48h, the ratio of transfection was tested by RT-PCR. The cell cycle and apoptosis were detected by flow cytometry. The expression of mRNA of cyclin E gene and A1 gene were detected by RT-PCR.RESULTS: After the transfection was finished for 48h, the number of lymphocyte in the G1-phase was increased, in the s-phase was depressed and in the G2 -phase was constant. The expression of cyclin E gene was lower, cyclin A1 gene was invaried.CONCLUSION: Cell cycle was inhibited in mouse mixed lymphocyte culture test and apoptosis was increasing when ADAR1 was restrained by the specific siRNA.