Effects Of Genistein On Host And HCMV Encoding Of IE Gene - Related MicroRNA Expression

Objective: 1 Genistein(Gen) was used as a drug to investigate the effect of Gen on human cytomegalovirus(HCMV) IE genes and acetyl-histone H3 binding major immediate early promoter(MIEP) in the human astrocyte(AS) infection by HCMV. 2 To investigate the effect of host’s and human HCMV’s micro RNA(mi RNA) on inhibiting HCMV infection in ASs.Methods: 1 ASs was derived from differentiation of human neural stem cell(NSC).HCMV proliferated in human embryonic lung fibroblast(HELFs). TCID50 was used to test the viral titer. 2 Cell counting kit-8(CCK-8) assay was used to test the cell proliferation and cytotoxicity with using different concentration of drug in order to screen out the appropriate concentration. 3 RT-q PCR was used to detect the expression changes of viral IE1,IE2 genes, host’s hsami R-200 mi RNA cluster and viral mi R-UL112. 4 Chromatin immunoprecipitation assay(Ch IP) was used to detect the change of acetylation of histone H3 binding MIEP.Results: 1 CCK-8 assay results showed that the cck-8 value of 10 μmol/L G+V group was higher than V group in the same time group. 2 Morphological observation showed that compared with control group, G group showed no significant difference. V group appeared significant cytopathic effect(CPE). The cell state in V+G group and G+V group were better than V group. And the cell state of G+V group which added drug first was better than V+G group which added drug last. 3 RT-q PCR results showed that compared with V group,10 μmol/L G+V and V+G group can reduce the IE1,IE2 genes expressions. 4 RT-q PCR results showed that 10 μmol/L G+V and V+G group can reverse the downregulation of hsa-mi R-200 mi RNA and mi R-UL112 when they contrasted with HCMV group.5 Ch IP results showed that compared with V group, 10 μmol/L G+V and V+G group can reduce acetylation of histone H3 binding MIEP.Conclusion: 1 10 μmol/L Gen can inhibit the AS’s abnormal proliferation caused by HCMV infection to some extent. 2 Gen can inhibit the expression of IE1,IE2 genes and reduce acetylation of histone H3 of MIEP to some extent, and reverse the down-regulation of partial host’s hsa-mi R-200 mi RNA cluster and viral mi R-UL112, maybe by which Gen play a role in inhibiting the IE protein.