UHRF1对前列腺癌PC-3细胞生物学特性的影响及相关机制

The Effects of UHRF1 on the Biological Characteristics of Prostate Cancer PC-3 Cells and Underlying Regulatory Mechanism

为了探讨泛素样含植物同源结构域和环指结构域1蛋白(ubiquitin-like with PHD and ring finger domains 1,UHRF1)对前列腺癌PC-3细胞生物学功能的影响及相关机制,我们用含有UHRF1和shRNA-UHRF1的慢病毒颗粒分别感染PC-3细胞,构建UHRF1沉默或过表达的稳转细胞株模型。利用实时荧光定量PCR(RT-qPCR)、蛋白免疫印迹(Western blot)和细胞免疫荧光实验验证感染后UHRF1表达水平和检测UHRF1对DNMT1、p16^(INK4A)的mRNA及蛋白表达水平的影响。CCK-8实验、划痕实验、Transwell实验和流式细胞术实验研究UHRF1对PC-3细胞增殖、侵袭迁移和周期凋亡的影响。结果显示,过表达UHRF1能够显著促进前列腺癌细胞的增殖、侵袭和迁移能力,并抑制细胞凋亡,调控细胞处于G2/M期,并且上调DNMT1表达水平,下调p16^(INK4A)表达水平,而沉默UHRF1的表达则作用相反。总之,UHRF1可能通过抑制p16^(INK4A)的表达促进前列腺癌细胞的增殖、侵袭和迁移,抑制细胞凋亡,UHRF1有可能成为前列腺癌潜在的治疗靶点。

To investigate the effects of ubiquitin-like with PHD and ring finger domains 1(UHRF1)on the cellular biology of prostate cancer PC-3 cells and underlying regulatory mechanisms,we transfected PC-3 cells with lentiviruses containing UHRF1 and shRNA-UHRF1 respectively,to construct stable transfected cell lines of UHRF1 silencing or overexpression.The expression level of UHRF1 and the effects of UHRF1 on DNMT1 and p16^(INK4A) expression at mRNA and protein levels were detected by real-time PCR(RT-qPCR),Western blot and immunofluorescence experiments,respectively.The effects of UHRF1 on the proliferation,migration,invasion,apoptosis and cell cycle distribution of PC-3 cells were studied by CCK-8,scratch assay,Transwell experiment and flow cytometry,respectively.Our data showed that overexpression of UHRF1 promoted cell proliferation,invasion and migration in PC-3 cells,inhibited cell apoptosis,regulated cells in G2/M phase,increased DNMT1 and decreased p16^(INK4A) expression at mRNA and protein levels.Downregulation of UHRF1 produces the opposite results.In summary,our results suggested that UHRF1 promote proliferation and migration but inhibits apoptosis of prostate cancer cells,likely by downregulating p16^(INK4A).UHRF1 may be a potential therapeutic target for prostate cancer.