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目的:探索卡波济肉瘤相关疱疹病毒(Kaposi’s sarcoma-associated herpesvirus,KSHV)体外感染人牙龈成纤维细胞(human gingival fibroblast,HGF)的途径和方法,为研究KSHV导致口腔卡波济肉瘤病变的机制提供依据。方法:用12-O-十四烷酰佛波醇-13-乙酯(12-O-tetradecanoylphorbol-13-acetate,TPA)刺激人原发性渗出性淋巴瘤(primary effusion lymphoma,PEL)细胞系的BC-3细胞,收集细胞上清(含KSHV病毒颗粒),感染HGF细胞。观察细胞病变效应(cytopatric effect,CPE),采用RT-PCR和Western blot分别检测HGF细胞内KSHV编码基因的转录情况和蛋白表达水平。结果:KSHV感染HGF细胞后可出现CPE;感染后采用RT-PCR可检测到不同时间点ORF26和ORF73基因的转录;感染后12 h可检测到vIL-6蛋白的表达。结论:KSHV可感染HGF细胞并建立潜伏感染,为进一步研究KSHV导致的口腔KS发病机制提供了较好的体外模型。
Objective: To explore the pathways and methods of in vitro infection of human gingival fibroblast (HGF) by Kaposi’s sarcoma-associated herpesvirus (KSHV). To investigate the mechanism of KSHV in the pathogenesis of oral Kaposi’s sarcoma Provide evidence. Methods: Human primary effusion lymphoma (PEL) cells were stimulated with 12-O-tetradecanoylphorbol-13-acetate (TPA) Line of BC-3 cells, collecting the cell supernatant (containing KSHV virus particles), infected HGF cells. The cytopatric effect (CPE) was observed. The transcription and expression of KSHV gene in HGF cells were detected by RT-PCR and Western blot respectively. Results: CPE could be detected in KSHV infected HGF cells. RT-PCR was used to detect the transcription of ORF26 and ORF73 genes at different time points after infection with KSHV. The expression of vIL-6 protein was detected 12 h after infection. CONCLUSIONS: KSHV can infect HGF cells and establish latent infection, which provides a good in vitro model for studying the pathogenesis of oral KS induced by KSHV.