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磷酸葡萄糖异构酶/自分泌运动因子的沉默降低了U87人胶质母细胞瘤细胞的迁移能力。

Silencing of phosphoglucose isomerase/autocrine motility factor decreases U87 human glioblastoma cell migration.

作者信息

Li Yang, Wei Zhenqing, Dong Bin, Lian Zhigang, Xu Yinghui

机构信息

Department of Neurosurgery, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, P.R. China.

出版信息

Int J Mol Med. 2016 Apr;37(4):998-1004. doi: 10.3892/ijmm.2016.2500. Epub 2016 Feb 23.

Abstract

Phosphoglucose isomerase/autocrine motility factor (PGI/AMF) is secreted by tumors and influences tumor growth and metastasis. In order to investigate the effects of silencing PGI/AMF on the migration and the sphere forming abilities of human glioblastoma U87 cells, as well as on the side population cells (SPCs), PGI/AMF was silenced using siRNA. Western blot analysis and RT-qPCR were used to assess the expression of PGI/AMF, Akt and SRY (sex determining region Y)-box 2 (SOX2). Wound healing, migration and tumorsphere formation assays were performed to assess invasion and metastatic potential. The proportion of SPCs was determined using Hoechst 33342 dye and flow cytometric analysis. PGI/AMF silencing inhibited the wound healing capacity and migration ability of U87 cells by 52.6 and 80.4%, respectively, compared with the scrambled siRNA (both P<0.001). Silencing of PGI/AMF decreased the proportion of SPCs in the U87 cells by 80.9% (P<0.01). The silencing of PGI/AMF decreased the number and size of tumorspheres by 53.1 and 39.9%, respectively, compared with the scrambled siRNA (both P<0.01). The silencing of PGI/AMF decreased the levels of phosphorylated Akt (-71.9%, P<0.001) compared with the scrambled siRNA, as well as the levels of the stemness marker, SOX2 (-61.7%, P<0.01). Taken together, these findings suggest that PGI/AMF silencing decreases migration, tumorsphere formation as well as the proportion of SPCs in glioblastoma U87 cells. We suggest that the Akt pathway is involved, and our results provide a potential new target for the treatment of glioblastoma.

摘要

磷酸葡萄糖异构酶/自分泌运动因子(PGI/AMF)由肿瘤分泌,并影响肿瘤的生长和转移。为了研究沉默PGI/AMF对人胶质母细胞瘤U87细胞迁移和球形成能力以及对侧群细胞(SPCs)的影响,使用小干扰RNA(siRNA)沉默PGI/AMF。采用蛋白质免疫印迹分析和逆转录定量聚合酶链反应(RT-qPCR)评估PGI/AMF、蛋白激酶B(Akt)和Y染色体性别决定区高迁移率族蛋白盒2(SRY-box2,SOX2)的表达。进行伤口愈合、迁移和肿瘤球形成试验以评估侵袭和转移潜能。使用Hoechst 33342染料和流式细胞术分析确定SPCs的比例。与乱序siRNA相比,沉默PGI/AMF分别使U87细胞的伤口愈合能力和迁移能力降低了52.6%和80.4%(均P<0.001)。沉默PGI/AMF使U87细胞中SPCs的比例降低了80.9%(P<0.01)。与乱序siRNA相比,沉默PGI/AMF使肿瘤球的数量和大小分别减少了53.1%和39.9%(均P<0.01)。与乱序siRNA相比,沉默PGI/AMF使磷酸化Akt水平降低了71.9%(P<0.001),同时使干性标志物SOX2水平降低了61.7%(P<0.01)。综上所述,这些发现表明沉默PGI/AMF可降低胶质母细胞瘤U87细胞的迁移、肿瘤球形成以及SPCs的比例。我们认为Akt信号通路参与其中,我们的结果为胶质母细胞瘤的治疗提供了一个潜在的新靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d74/4790702/792d97655ca0/IJMM-37-04-0998-g00.jpg

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