抑制素对HCV复制的影响

发布时间：2019-03-21 11:27

【摘要】：目的研究抑制素(PHB)对HCV复制的影响。方法全长基因组HCV RNA体外转染人肝癌细胞系Huh7.5,构建HCV全基因组细胞模型,分别收集24、48、72、96 h培养上清液检测HCV拷贝数;间接免疫荧光实验检测HCV核心蛋白的表达;透射电镜观察HCV感染细胞Huh 7.5-HCV的超微结构改变,鉴定HCV全基因组细胞模型。采用实时定量PCR、Western Blotting方法研究PHB在Huh 7.5-HCV细胞中的表达情况。应用RNA干扰技术检测PHB对HCV RNA的复制情况。两组间比较采用t检验。结果鉴定结果显示,成功构建HCV全基因组细胞模型。HCV RNA转染Huh7.5细胞24和48 h后,PHB mRNA表达水平分别是13.41±1.35和16.45±1.76,与对照组(1.01±0.57和1.01±0.87)相比差异均有统计学意义(t值分别为29.540、31.361,P值均0.01);PHB蛋白表达水平亦显著高于对照组(P值均0.01)。PHB的RNA干扰质粒(shRNA-PHB)转染Huh7.5-HCV细胞24和48h后,HCV RNA的水平分别为64.32±5.49和84.45±7.06,显著高于对照组(shRNA-control组,10.52±1.57和16.34±2.97,t值分别为29.538、25.908,P值均0.01)。结论 Huh7.5-HCV细胞中PHB表达的升高可能与HCV RNA的感染有关,PHB蛋白对HCV RNA复制有一定的抑制作用。
[Abstract]:Objective To study the effect of inhibin (PHB) on the replication of HCV. Methods The whole-length genomic HCV RNA was transfected into human liver cancer cell line Huh7.5, and the whole genome cell model of HCV was constructed,24,48,72 and 96 hours of culture supernatant were collected to test the copy number of HCV. The indirect immunofluorescence assay was used to detect the expression of HCV core protein. The ultrastructure of HCV-infected cells Huh 7.5-HCV was observed by transmission electron microscope, and the whole-genome cell model of HCV was identified. The expression of PHB in Huh 7.5-HCV cells was studied by real-time quantitative PCR and Western Blotting. The replication of HCV RNA was detected by RNA interference technique. A t-test was used between the two groups. Results The results showed that the whole genome model of HCV was successfully constructed. The expression of PHB mRNA was 13.41-1.35 and 16.45-1.76, respectively, compared with the control group (1.01, 0.57 and 1.01-0.87), and the expression of PHB was significantly higher than that in the control group (P <0.01). After 24 and 48 h, the level of HCV RNA was 64.32, 5.49 and 84.45-7.06, which was significantly higher than that in control group (shRNA-control group, 10.52-1.57 and 16.34-2.97, t-value was 29.538, 25.908, P <0.01). Conclusion The increase of PHB expression in Huh7.5-HCV cells may be related to the infection of HCV RNA, and PHB protein can inhibit the replication of HCV RNA.
【作者单位】：西安交通大学第二附属医院感染科;
【基金】：国家自然科学基金资助项目(81170393)
【分类号】：R512.63

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