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    张庆, 秦苏萍, 李小翠, 王晓天, 刘晓梅, 周峰. 细胞周期蛋白激酶抑制调控蛋白p21在HHV-8病毒裂解复制周期的作用[J]. 徐州医科大学学报, 2024, 44(2): 95-99. DOI: 10.3969/j.issn.2096-3882.2024.02.003
    引用本文: 张庆, 秦苏萍, 李小翠, 王晓天, 刘晓梅, 周峰. 细胞周期蛋白激酶抑制调控蛋白p21在HHV-8病毒裂解复制周期的作用[J]. 徐州医科大学学报, 2024, 44(2): 95-99. DOI: 10.3969/j.issn.2096-3882.2024.02.003
    shu
    ZHANG Qing, QIN Suping, LI Xiaocui, WANG Xiaotian, LIU Xiaomei, ZHOU Feng. Effect of cyclin-dependent kinase inhibitor p21 on regulating viral lytic replication of human herpesvirus 8[J]. Journal of Xuzhou Medical University, 2024, 44(2): 95-99. DOI: 10.3969/j.issn.2096-3882.2024.02.003
    Citation: ZHANG Qing, QIN Suping, LI Xiaocui, WANG Xiaotian, LIU Xiaomei, ZHOU Feng. Effect of cyclin-dependent kinase inhibitor p21 on regulating viral lytic replication of human herpesvirus 8[J]. Journal of Xuzhou Medical University, 2024, 44(2): 95-99. DOI: 10.3969/j.issn.2096-3882.2024.02.003
    shu

    细胞周期蛋白激酶抑制调控蛋白p21在HHV-8病毒裂解复制周期的作用

    Effect of cyclin-dependent kinase inhibitor p21 on regulating viral lytic replication of human herpesvirus 8

      摘要
    • 摘要: 目的 研究细胞周期蛋白激酶抑制调控蛋白p21对人类疱疹病毒8型(human herpesvirus 8,HHV-8)病毒裂解复制周期的影响。方法 组蛋白去乙酰化酶(histone deacetylase, HDAC)抑制剂SAHA预处理后,倒置荧光显微镜观察红色荧光蛋白(RFP)阳性的iSLK.219细胞数,实时定量PCR检测TREx-K-Rta BCBL-1细胞中HHV-8病毒相关基因的mRNA水平。脂质体转染p21-siRNA后,免疫印迹法检测iSLK.219和TREx-K-Rta BCBL-1细胞中p21蛋白表达,计算RFP阳性iSLK.219细胞百分率,检测TREx-K-Rta BCBL-1细胞中ORF50和PAN的mRNA水平,CCK-8法和台盼蓝染色观察细胞存活情况。结果 SAHA显著增强iSLK.219细胞RFP阳性率、TREx-K-Rta BCBL-1细胞中HHV-8裂解复制周期相关基因ORF50、PAN及K8.1的mRNA水平和p21蛋白表达,差异有统计学意义(P<0.05)。siRNA沉默 p21后,iSLK.219细胞RFP阳性率、TREx-K-Rta BCBL-1细胞中HHV-8裂解复制周期相关基因ORF50和PAN mRNA水平显著下降,差异有统计学意义(P<0.05),且保护SAHA介导的TREx-K-Rta BCBL-1细胞死亡。结论 抑制 HDAC活性通过调控p21促进HHV-8病毒裂解复制。

       

      Abstract: 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.

       

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