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PUM2通过抑制BTG1表达促进胶质母细胞瘤细胞增殖和迁移。

PUM2 Promotes Glioblastoma Cell Proliferation and Migration via Repressing BTG1 Expression.

作者信息

Wang Yuanyu, Sun Weili, Yang Jiankai, Yang Liang, Li Chen, Liu Hongjiang, Liu Xiaopeng, Jiao Baohua

机构信息

Department of Neurosurgery, the Second Hospital of Hebei Medical University.

出版信息

Cell Struct Funct. 2019;44(1):29-39. doi: 10.1247/csf.18030.

DOI:10.1247/csf.18030
PMID:30787206
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11926404/
Abstract

PUM2, an RNA binding protein, is known to promote stem cell proliferation via repressing expressions of cell cycle genes. Similar with stem cells, malignant cells are characterized by unlimited proliferation and remote migration. However, roles of PUM2 in cancer development are controversial. Here, we investigated PUM2's role in glioblastoma development and its relationship with the cell cycle regulator BTG1. Immunoblotting and RT-qPCR were used to evaluate protein expression level and transcript level, respectively. ShRNAs were designed to knock down PUM2 and BTG1 expression. CCK-8 assay was used to evaluate cell viability. Cell migration assay and evasion assay were used to evaluate metastatic capability of glioblastoma cell. RNA pull-down assay and RNA immunoprecipitation assay were used to test the interaction between PUM2 and BTG1 3'UTR. PUM2 expression is elevated in glioblastoma tumor tissues as well as glioblastoma cell lines. PUM2 knockdown remarkably suppresses glioblastoma cell proliferation and migration. In addition, PUM2 knockdown increases BTG1 expression. RNA pull-down assay and RNA immunoprecipitation assay show PUM2 binds to BTG1 3'UTR directly. Furthermore, knockdown of BTG1 reverses the effect of PUM2 knockdown on glioblastoma cell proliferation and migration. Our results suggest that PUM2 promote glioblastoma development via repressing BTG1 expression.Key words: PUM2, BTG1, glioblastoma, cell proliferation, metastasis.

摘要

PUM2是一种RNA结合蛋白，已知其通过抑制细胞周期基因的表达来促进干细胞增殖。与干细胞相似，恶性细胞的特征是无限增殖和远距离迁移。然而，PUM2在癌症发展中的作用存在争议。在此，我们研究了PUM2在胶质母细胞瘤发展中的作用及其与细胞周期调节因子BTG1的关系。分别使用免疫印迹和RT-qPCR来评估蛋白表达水平和转录水平。设计短发夹RNA（shRNAs）以敲低PUM2和BTG1的表达。使用CCK-8测定法评估细胞活力。使用细胞迁移测定法和侵袭测定法评估胶质母细胞瘤细胞的转移能力。使用RNA下拉测定法和RNA免疫沉淀测定法测试PUM2与BTG1 3'非翻译区（UTR）之间的相互作用。PUM2在胶质母细胞瘤肿瘤组织以及胶质母细胞瘤细胞系中的表达升高。敲低PUM2可显著抑制胶质母细胞瘤细胞的增殖和迁移。此外，敲低PUM2可增加BTG1的表达。RNA下拉测定法和RNA免疫沉淀测定法表明PUM2直接与BTG1 3'UTR结合。此外，敲低BTG1可逆转敲低PUM2对胶质母细胞瘤细胞增殖和迁移的影响。我们的结果表明，PUM2通过抑制BTG1的表达促进胶质母细胞瘤的发展。关键词：PUM2；BTG1；胶质母细胞瘤；细胞增殖；转移

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9304/11926404/d423c3e5f96e/csf_44_18030-f007.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9304/11926404/d423c3e5f96e/csf_44_18030-f007.jpg

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