Cui Ying, Li Jiang, Zheng Fei, Ouyang Yongri, Chen Xi, Zhang Lei, Chen Yang, Wang Lin, Mu Shijie, Zhang Huizhong
Department of Blood Transfusion, Tangdu Hospital, The Fourth Military Medical University, Xi'an, Shanxi, China (mainland).
Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi'an, Shanxi, China (mainland).
Med Sci Monit. 2016 Feb 5;22:380-6. doi: 10.12659/msm.895312.
BACKGROUND The epithelial-mesenchymal transition (EMT) has been shown to be involved in the process of invasion and metastasis of prostate cancer. SIRT1 is the mammalian homologue of the silent information regulator 2 (Sir2) gene, and is abnormally expressed in prostate cancer cells. Therefore, it is hypothesized that SIRT1 mediates the invasion/metastatic ability of prostate cancer via EMT regulation. This study thus investigated the effect of SIRT1 gene on the invasion and migration of prostate cancer cell line PC-3 via the small interference RNA (siRNA) against SIRT1. MATERIAL AND METHODS SiRNA construct was transfected into PC-3 cells, which were tested for the cell migration and invasion ability by scratch assay and Transwell migration assay, respectively. Expression levels of vimentin, E-cadherin, and N-cadherin were further quantified by Western blotting and RT-PCR. RESULTS Both mRNA and protein levels of SIRT1 were depressed after siRNA transfection, along with weakened migration and invasion ability of PC-3 cells. Elevated E-cadherin and suppressed N-cadherin and vimentin were observed in those transfected cells. CONCLUSIONS The silencing of SIRT1 gene in PC-3 cells can suppress the movement, migration, and invasion functions of prostate cancer cells, possibly via the down-regulation of mesenchymal markers vimentin and N-cadherin accompanied with up-regulation of epithelial marker N-cadherin, thus reversing the EMT process.
背景 上皮-间质转化(EMT)已被证明参与前列腺癌的侵袭和转移过程。SIRT1是沉默信息调节因子2(Sir2)基因的哺乳动物同源物,在前列腺癌细胞中异常表达。因此,推测SIRT1通过EMT调节介导前列腺癌的侵袭/转移能力。本研究通过针对SIRT1的小干扰RNA(siRNA)研究SIRT1基因对前列腺癌细胞系PC-3侵袭和迁移的影响。
材料与方法 将siRNA构建体转染到PC-3细胞中,分别通过划痕试验和Transwell迁移试验检测细胞的迁移和侵袭能力。通过蛋白质印迹和RT-PCR进一步定量波形蛋白、E-钙黏蛋白和N-钙黏蛋白的表达水平。
结果 siRNA转染后,SIRT1的mRNA和蛋白水平均降低,同时PC-3细胞的迁移和侵袭能力减弱。在这些转染细胞中观察到E-钙黏蛋白升高,N-钙黏蛋白和波形蛋白受到抑制。
结论 PC-3细胞中SIRT1基因的沉默可抑制前列腺癌细胞的运动、迁移和侵袭功能,可能是通过下调间质标志物波形蛋白和N-钙黏蛋白,同时上调上皮标志物E-钙黏蛋白,从而逆转EMT过程。