Objective To explore the role of cyclin-dependent kinase inhibitor p21 in regulating the lytic replication of human herpesvirus 8(KSHV).Methods After pretreatment with the histone deacetylase(HDAC)inhibitor SAHA,the number of red fluorescent protein(RFP)-positive iSLK.219 cells was observed by inverted fluorescence microscopy.The mRNA levels of HHV-8 virus-related genes in TREx-K-Rta BCBL-1 cells were detected by real-time quantitative PCR.After liposomal transfection of p21-siRNA,the expression of p21 protein in iSLK.219 and TREx-K-Rta BCBL-1 cells was detected by Western blot;the percentage of RFP-positive iSLK.219 cells was calculated,and the levels of ORF50 and PAN mRNA in TREx-K-Rta BCBL-1 cells were detected,and the survival of iSLK.219 and TREx-K-Rta BCBL-1 cells were measured by CCK-8 assay and Trypan blue staining.Results SAHA dramatically enhanced the percent of RFP-positive iSLK.219 cells,and the mRNA levels of ORF50,PAN and K8.1,and p21 expression(P＜0.05).Knockdown of p21 significantly suppressed the percentage of RFP-positive iSLK.219 cells,and the mRNA levels of ORF50 and PAN(P＜0.05),and protected SAHA-mediated cell death in BCBL-1 cells.Conclusions Inhibition of HDAC activity promotes HHV-8 replication through regulating p21.
关键词
人类疱疹病毒8型/病毒裂解复制周期/组蛋白去乙酰化酶/细胞周期蛋白激酶抑制调控蛋白p21/细胞死亡
Key words
human herpesvirus 8/viral lytic replication/histone deacetylase/cyclin-dependent kinase inhibitor p21/cell death
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