α27基因特异性siRNA对单纯疱疹病毒Ⅱ型感染宿主细胞影响.docVIP

α27基因特异性siRNA对单纯疱疹病毒Ⅱ型感染宿主细胞影响 　　[摘要] 目的 探讨在RNA干扰技术条件上，干扰α27基因表达对单纯疱疹病毒Ⅱ型（HSV-Ⅱ）宿主细胞的影响。 方法 体外合成四对α27基因特异性siRNA（R1、R2、R3、R4组）和阳性、阴性对照siRNA，用Lipofectamine2000转染Vero细胞后接种HSV-Ⅱ，感染后用MTT比色法分别于12、24、36、48、60、72 h测定细胞的存活情况。 结果 转染了特异性siRNA的细胞OD值较空白组高，细胞存活情况较好，其中R2和R4组存活情况更好。结论 α27基因特异性siRNA对宿主细胞具有保护作用。按照同样设计原则合成的不同序列siRNA，其抑制效果存在明显差异。 　　[关键词] α27基因；siRNA；RNA干扰；单纯疱疹病毒Ⅱ型；Vero细胞 　　[中图分类号] Q936 [文献标识码] A [文章编号] 1674-4721（2015）05（b）-0007-04 　　Effect of α27-specific siRNA on host cells infecting by herpes simplex virus type Ⅱ 　　YANG Zhao-lin CHEN Yin WANG Li ZHAO Shao-feng▲ SHAO Yan-ling 　　Department of Gynaecology and Obstetrics，Guangfo Hospital Affiliated to the Guangzhou University of Medical Science the Second Peoples Hospital of Nanhai District in Foshan City，Guangdong Province，Foshan 528251，China 　　[Abstract] Objective To determine the effect of α27-specific siRNA on host cells infecting by herpes simplex virus type Ⅱ （HSV-Ⅱ）. Methods Four α27-specific siRNAs （R1，R2，R3，R4 group） and the positive，negative control siRNAs were synthesized in vitro by chemical process.The Vero cells were transfected with siRNAs using Lipofectamine2000 followed by HSV-Ⅱinfecting.The activity of the cells were measured at 12，24，36，48，60，72 h by MTT. Results Groups with α27-specific siRNA had higher OD values by MTT colorimetric assay than blank cells，with better survive situation.R2 and R4 group had better survival than other groups. Conclusion α27-specific siRNA can protect host cells of HSV-Ⅱ.There are obvious differences of inhibition effect of siRNA which are synthesized in accordance with the same priciples. 　　[Key words] α27 gene；siRNA；RNAi；HSV-Ⅱ；Vero cell 　　生殖器疱疹（genital herpes，GH）是由单纯疱疹病毒（herpes simplex virus，HSV）感染生殖器和肛门部位皮肤、黏膜而引起的一种慢性、易复发的性传播疾病。GH的病原体90%为HSV-Ⅱ型[1]。HSV-Ⅱ基因组为一线性双链DNA分子，根据基因表达时序的不同，分为立即早期、早期、晚期基因，分别称为α、β、γ基因，分别编码立即早期蛋白、早期蛋白及晚期蛋白。立即早期基因的转录出现在病毒DNA复制之前，调节β、γ基因的表达。迄今为止，人们已知的HSV编码的立即早期蛋白有5个，它们被称为感染细胞多肽分子（infected cell polypeptides，ICPs），包括ICP0、ICP4、ICP22、ICP27和ICP47[