Li Ying, Xu Huiling, Tan Xiaohua, Cui Qinghua, Gu Wenyi, Pan Zemin, Yang Lei, Wu Shuyuan, Wang Xiaolu, Li Dongmei
Key Laboratory of Xinjiang Endemic and Ethnic Diseases/NHC Key Laboratory of Prevention and Treatment of Central Asia High Incidence Diseases, Shihezi University, Shihezi, 832002, Xinjiang, China.
School of Medicine, Tarim University, Alaer, 843300, Xinjiang, China.
Arch Virol. 2023 Jan 7;168(2):39. doi: 10.1007/s00705-022-05626-0.
The disease caused by Kaposi's sarcoma-associated herpesvirus (KSHV) is one of the major causes of death of individuals with acquired immunodeficiency syndrome (AIDS). Development of anti-KSHV drugs is thus crucial. In this study, we investigated the effect of parthenolide (PTL) on the proliferation and NF-κB signaling pathway of KSHV-infected cells. iSLK.219 and KSHV-infected SH-SY5Y cells (SK-RG) were treated with PTL, TaqMan real-time quantitative PCR was used to determine the number of copies of the KSHV genome, and mRNA and protein expression of KSHV genes were analyzed by real-time PCR and immunocytochemistry. A cell viability test was used to measure cell proliferation, and flow cytometry was used to examine the effect of the drug on the cell cycle. Cyclin D1, CDK6, CDK4, and NF-κB-related proteins, including IKKβ, P-p65, and P-IKB-α, were detected by Western blot. The results showed that PTL altered the morphology of the cells, reduced the KSHV copy number, and suppressed the production of ORF50, K8.1, and v-GPCR mRNA and the LANA, ORF50, and K8.1 proteins. It blocked the G1 phase in iSLK.219 cells and decreased the levels of cyclin D1, CDK6, and CDK4 as well as the levels of NF-κB signaling proteins, including IKKβ, P-p65, and P-IKB-α. Together, these results suggest that PTL is a candidate drug that can decrease KSHV pathogenicity by suppressing cell proliferation and inhibiting the NF-κB signaling pathway in KSHV-infected cells.
卡波西肉瘤相关疱疹病毒（KSHV）引起的疾病是获得性免疫缺陷综合征（AIDS）患者的主要死因之一。因此，开发抗KSHV药物至关重要。在本研究中，我们研究了小白菊内酯（PTL）对KSHV感染细胞增殖和NF-κB信号通路的影响。用PTL处理iSLK.219和KSHV感染的SH-SY5Y细胞（SK-RG），采用TaqMan实时定量PCR测定KSHV基因组的拷贝数，并通过实时PCR和免疫细胞化学分析KSHV基因的mRNA和蛋白表达。采用细胞活力试验检测细胞增殖，用流式细胞术检测药物对细胞周期的影响。通过蛋白质免疫印迹法检测细胞周期蛋白D1、细胞周期蛋白依赖性激酶6（CDK6）、细胞周期蛋白依赖性激酶4（CDK4）以及NF-κB相关蛋白，包括IκB激酶β（IKKβ）、磷酸化p65（P-p65）和磷酸化IκB-α（P-IKB-α）。结果表明，PTL改变了细胞形态，降低了KSHV拷贝数，抑制了开放阅读框50（ORF50）、K8.1和病毒G蛋白偶联受体（v-GPCR）mRNA的产生以及潜伏核抗原（LANA）蛋白、ORF50蛋白和K8.1蛋白的产生。它阻断了iSLK.219细胞的G1期，并降低了细胞周期蛋白D1、CDK6和CDK4的水平以及NF-κB信号蛋白的水平，包括IKKβ、P-p65和P-IKB-α。综上所述，这些结果表明PTL是一种候选药物，可通过抑制细胞增殖和抑制KSHV感染细胞中的NF-κB信号通路来降低KSHV致病性。
