Xu Chenyang, Li Zhenjiang, He Tao, Yuan Bingjian, Ding Bingqian
Department of Neurosurgery, Huaihe Hospital of Henan University No. 1, North Baogong Rd, Gulou Kaifeng Henan 475000 China
RSC Adv. 2019 Apr 8;9(19):10870-10880. doi: 10.1039/c8ra09546a. eCollection 2019 Apr 3.
Previous studies have found that long noncoding RNA (lncRNA) protein tyrosine phosphatase, receptor type, G, antisense (PTPRG-AS1) was upregulated in glioma cells. Our study aimed to explore the detailed molecular mechanisms of PTPRG-AS1 involved in glioma progression. qRT-PCR assay was performed to measure the expressions of PTPRG-AS1 and microRNA-185-5p (miR-185-5p). Cell viability, migration, invasion, and apoptosis were determined by CCK-8 assay, colony formation assay, transwell assay, and flow cytometry assay. Autophagy was evaluated using GFP-LC3 puncta analysis and western blot. Luciferase reporter and RIP assays were employed to explore the association between PTPRG-AS1 and miR-185-5p. Our data showed PTPRG-AS1 was upregulated in glioma cells and tissues. Besides, high expression of PTPRG-AS1 was positively associated with a low survival rate. Upregulation of PTPRG-AS1 promoted proliferation, migration, invasion, colony formations, and autophagy, and inhibited cell apoptosis in U373-MG cells. By contrast, PTPRG-AS1 downregulation had the inverse effect in SHG44 cells. PTPRG-AS1 negatively regulated the expression of miR-185-5p in U373-MG and SHG44 cells and the expression of miR-185-5p was decreased in glioma tissues and cells. In addition, miR-185-5p overexpression suppressed proliferation, metastasis, colony formations, and autophagy, while inducing cell apoptosis in SHG44 cells. As expected, miR-185-5p depletion exhibited the inverse effect in U373-MG cells. Enhanced expression of miR-185-5p attenuated the effect of PTPRG-AS1 upregulation on U373-MG cells, while silencing of miR-185-5p undermined the effect of downregulation of PTPRG-AS1 on SHG44 cells. Our data disclosed that LncRNA PTPRG-AS1 was upregulated in glioma cells and tissues. PTPRG-AS1 regulated glioma proliferation, invasion, migration, apoptosis and autophagy by sponging miR-185-5p . A new signaling pathway PTPRG-AS1/miR-185-5p was first observed in glioma.
先前的研究发现，长链非编码RNA（lncRNA）蛋白酪氨酸磷酸酶受体型G反义链（PTPRG-AS1）在胶质瘤细胞中表达上调。我们的研究旨在探索PTPRG-AS1参与胶质瘤进展的详细分子机制。采用qRT-PCR检测法检测PTPRG-AS1和微小RNA-185-5p（miR-185-5p）的表达。通过CCK-8检测法、集落形成检测法、Transwell检测法和流式细胞术检测法测定细胞活力、迁移、侵袭和凋亡情况。使用绿色荧光蛋白标记的微管相关蛋白1轻链3（GFP-LC3）斑点分析和蛋白质免疫印迹法评估自噬。采用荧光素酶报告基因检测和RNA免疫沉淀（RIP）检测来探究PTPRG-AS1与miR-185-5p之间的关系。我们的数据表明，PTPRG-AS1在胶质瘤细胞和组织中表达上调。此外，PTPRG-AS1的高表达与低生存率呈正相关。PTPRG-AS1的上调促进了U373-MG细胞的增殖、迁移、侵袭、集落形成和自噬，并抑制了细胞凋亡。相比之下，PTPRG-AS1的下调在SHG44细胞中产生相反的作用。PTPRG-AS1在U373-MG和SHG44细胞中负向调节miR-185-5p的表达，且miR-185-5p在胶质瘤组织和细胞中的表达降低。此外，miR-185-5p的过表达抑制了SHG44细胞的增殖、转移、集落形成和自噬，同时诱导细胞凋亡。正如预期的那样，miR-185-5p的缺失在U373-MG细胞中产生相反的作用。miR-185-5p表达的增强减弱了PTPRG-AS1上调对U373-MG细胞的影响，而miR-185-5p的沉默则削弱了PTPRG-AS1下调对SHG44细胞的影响。我们的数据表明，lncRNA PTPRG-AS1在胶质瘤细胞和组织中表达上调。PTPRG-AS1通过海绵吸附miR-185-5p来调节胶质瘤的增殖、侵袭、迁移、凋亡和自噬。在胶质瘤中首次观察到一条新的信号通路PTPRG-AS1/miR-185-5p。
