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

作　　者：张庆[1] 秦苏萍 李小翠 王晓天 刘晓梅 周峰 ZHANG Qing;QIN Suping;LI Xiaocui;WANG Xiaotian;LIU Xiaomei;ZHOU Feng(Department of Medical Examinafiom,Xuzhou Cancer Hospital,Xuzhou,Jiangsu 221009,China;Jiangsu Key Laboratory of Immunity and Metabolism,Xuzhou Medical University,Xuzhou,Jiangsu 221004)

机构地区：[1]徐州市肿瘤医院检验科,江苏徐州221009 [2]徐州医科大学江苏省免疫与代谢重点实验室,江苏徐州221004

出　　处：《徐州医科大学学报》2024年第2期95-99,共5页Journal of Xuzhou Medical University

基　　金：国家自然科学基金面上项目(81971179);江苏省教育厅项目(20KJA320004)。

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。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 RFPpositive 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 

分 类 号：R373[医药卫生—病原生物学]