HIV-1Vpr基因在肿瘤细胞中诱导G2期停滞、细胞致死效应及其核定位功能研究

Cell cycle G2 arrest,cell death and nuclear localization induced by human immunodeficiency virus type 1 protein R(Vpr) in cervical cancer cells

目的 通过研究HIV - 1Vpr基因在宫颈癌细胞HeLa中诱导细胞周期G2停滞、细胞致死效应及其核定位功能 ,探讨将Vpr用于肿瘤治疗的可能性。方法 用脂质体转染的方法 ,将携带Vpr或其突变体VprX基因的质粒转入HeLa细胞内 ,用流式细胞仪分析Vpr诱导的G2停滞和细胞致死性 ;用荧光显微镜观察GFP -Vpr融合蛋白的荧光确定其核定位功能。结果 Vpr具有核定位功能。Vpr和VprX蛋白都具有细胞致死性 ,但是VprX蛋白不能使细胞停滞到G2期。结论 首次报道了Vpr的G2期停滞和细胞致死功能在哺乳动物细胞中是互相独立的。

Objective To explore the possibility of using Vpr as a biological agent for anticancer therapy.Methods A HIV-1 wild type Vpr and a mutant vprX gene were expressed and tested in a cervical cancer HeLa cell line.The ability of Vpr to induce cell cycle G2 arrest and cell death was measured by flow cytometric analysis.Nuclear localization of Vpr was visualized by fluorescent light emitted from the green fluorescent protein (GFP)-Vpr fusion protein.Results Consistent with early reports,Vpr,not VprX,induced G2 arrest.However,both Vpr and VprX were able to induce cell death in HeLa cells.We also demonstrated that the nuclear localization of Vpr could be determined by using a GFP-Vpr fusion protein.Conclusion This is the first report documenting in mammalian cells that G2 arrest and cell death induced by Vpr are two independent functions.The unique biological properties of Vpr shown in cervical cancer cells suggest that Vpr may be a useful biological agent for anti-cancer therapy.