The online version of this article (https://doi.org/10.1186/s13046-017-0665-3) contains supplementary material, which is available to authorized users.
Golgi Membrane Protein 1 (GOLM1), a protein involved in the trafficking of proteins through the Golgi apparatus, has been shown to be oncogenic in a variety of human cancers. Here, we examined the role of GOLM1 in the development of human glioma.
qRT-PCR, immunohistochemistry, and western blot analysis were performed to evaluate GOLM1 levels in cell lines and a cohort of primary human glioma and non-neoplastic brain tissue samples. Glioma cell lines were modified with lentiviral constructs expressing short hairpin RNAs targeting GOLM1 or overexpressing the protein to assess function in proliferation, viability, and migration and invasion in vitro using EdU, CCK8, clone-forming, Transwell assays, 3D tumor spheroid invasion assay and in vivo in orthotopic implantations. Protein lysates were used to screen a membrane-based antibody array to identify kinases mediated by GOLM1. Specific inhibitors of PDGFRα (AG1296) and AKT (MK-2206) were used to examine the regulation of PDGFA/PDGFRα on GOLM1 and the underlying pathway respectively.
qRT-PCR, immunohistochemistry and western blot analysis revealed GOLM1 expression to be elevated in glioma tissues and cell lines. Silencing of GOLM1 attenuated proliferation, migration, and invasion of U251, A172 and P3#GBM (primary glioma) cells, while overexpression of GOLM1 enhanced malignant behavior of U87MG cells. We further demonstrated that activation of AKT is the driving force of GOLM1-promoted glioma progression. The last finding of this research belongs to the regulation of PDGFA/PDGFRα on GOLM1, while GOLM1 was also a key element of PDGFA/PDGFRα-mediated activation of AKT, as well as the progression of glioma cells.
PDGFA/PDGFRα-regulated GOLM1 promotes glioma progression possibly through activation of a key signaling kinase, AKT. GOLM1 interference may therefore provide a novel therapeutic target and improve the efficacy of glioma treatment, particularly in the case of the proneural molecular subtype of human glioma.
Additional file 1: Figure S1. Expression of GOLM1 were analyzed in normal brain tissues (n = 4), WHO II gliomas (n = 4) and WHO III-IV gliomas (n = 11). (TIFF 682 kb)
Additional file 2: Figure S2. (a-b) Representative images and graphic representation of colony forming assays for U251- and A172-NC or sh-GOLM1 cells. (c) Representative images and graphic representation of colony forming assays for U87MG- Lenti-NC or -Lenti-GOLM1 cells. Data are presented as the mean ± SEM. (TIFF 5314 kb)
Additional file 3: Figure S3. ImageJ was introduced to assess the western blot results in in Fig. 6c (a) and 6d (b). Data are presented as the mean ± SEM. (TIFF 2306 kb)
Additional file 4: Figure S4. (a) Kinases and genes downstream of AKT in P3#GBM cells were analyzed by western blot. (b) ImageJ was introduced to assess the western blot results in (a). Data are presented as the mean ± SEM. (TIFF 2694 kb)
Additional file 5: Table S1. Association of p-PDGFRα with GOLM1 protein levels in primary human GBM samples (n = 29). (DOCX 14 kb)
Additional file 6: Figure S5. ImageJ was introduced to assess the western blot results in in Fig. 8a (a), 8c (b), 8d (c) and 8i (d). Data are presented as the mean ± SEM. (TIFF 2057 kb)
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- PDGFA/PDGFRα-regulated GOLM1 promotes human glioma progression through activation of AKT
- BioMed Central
Journal of Experimental & Clinical Cancer Research
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