Journal: Frontiers in Oncology

Article Title: The role of tumor-associated macrophages in glioma cohort: through both traditional RNA sequencing and single cell RNA sequencing

doi: 10.3389/fonc.2023.1249448

Figure Lengend Snippet: (A) The cck8 assay in U87 cells; (B) The cck8 assay in U251 cells; (C) The Edu assay in U87 cells; (D) The Edu assay in U251 cells.

Article Snippet: Human glioma cell lines U251 and U87 were purchased from Beijing Zhongyuan Heju Biotechnology Co., Ltd. (Beijing, China).

Techniques: CCK-8 Assay, EdU Assay

Journal: International Journal of Molecular Sciences

Article Title: LPPR5 Expression in Glioma Affects Growth, Vascular Architecture, and Sunitinib Resistance

doi: 10.3390/ijms23063108

Figure Lengend Snippet: In-silico analysis of LPPR5 in physiological brain tissue and primary brain tumor. ( a ) LPPR5 expression is physiologically high in human adult brain. Outside the CNS, LPPR5 is compara-tively highly expressed in testis [GTeX gene expression data for LPPR5 in brain (blue, Hippocampus light green) and non-brain tissue (dark green). Expression values are shown in TPM (Tran-scripts Per Million), calculated from a gene model with isoforms collapsed to a single gene. No other normalization steps have been applied. Box plots are shown as median and 25th and 75th per-centiles; points are displayed as outliers if they are above or below 1.5 times the interquartile range]. ( b ) LPPR5 is significantly un-der-expressed in (high-grade) glioma. Relative under-expression rank is median rank across all analyses in gene expression in Sun Brain, and respectively Roth normal in Oncomine [p for medi-an-ranked analysis, Oncomine visualization and statistical anal-ysis]. ( c ) A subgroup of high-grade glioma has decreased LPPR5 expression; in lower grade glioma, the ratio of tumors with low LPPR5 expression is smaller [mRNA expression data from Sun Brain, each bar represents a single tumor, II and III indicates astrocytoma WHO grade]. ( d ) On the DNA-level, LPPR5 is deleted in most oligodendrogliomas and to a lesser degree in lower grade astrocytic gliomas and GBM [Oncomine visualization of LPPR5 DNA copy number units of the tumors in Kotliarov Brain, each bar represents a single tumor]. ( e ) LPPR5 expression is significantly lower in the mesenchymal expression subtype of GBM ( n = 49) compared to classical ( n = 39, * p = 0.0451) and proneural ( n = 29, **** p < 0.0001) subtypes. Expression in the neural subtype ( n = 26) is significantly lower than in proneural subtype (* p = 0.0162) [TCGA via cBio]. ( f ) Histogram plot of 70 human Glioma cell lines screened, 50 show low expression of LPPR5 [Cancer Cell Line Encyclopedia, Broad Institute].

Article Snippet: Moreover, in a screening of 70 human glioma cell lines from the Cancer Cell Line Encyclopedia (Broad Institute), only 4 cell lines showed a significant expression of LPPR5 (LN319, KNS42, TM31, and NMCG1; f).

Techniques: In Silico, Expressing, Gene Expression

Journal: BMC Cancer

Article Title: Valproic acid-induced amphiregulin secretion confers resistance to temozolomide treatment in human glioma cells

doi: 10.1186/s12885-019-5843-6

Figure Lengend Snippet: VPA regulated the cytotoxic effects of TMZ in U87MG cells. Cells were treated with VPA (500 μM) or TMZ (500 μM), alone or in combination, for 48 h. a Cell cytotoxicity was measured by an XTT-based assay. Data are presented as mean ± S.E.M. of at least 3 independent experiments. * P < 0.05, compared with the control (0 μM). # P < 0.05, compared with cells treated with TMZ (500 μM) alone. b Cell cycle was analyzed by PI staining and flow cytometry. Sub-G1, G0/G1, S and G2/M indicate different cell cycle phases (Upper panel). Bar graphs present the percentage of the number of cells in each phase. Data are presented as mean ± S.E.M. from 3 independent experiments (Lower panel). c-e The effects of VPA on TMZ-induced apoptosis and autophagy were assessed by Western blot using cleaved PARP, cleaved caspase-3, and LC3B antibodies

Article Snippet: The human glioblastoma cell line U87MG, DBTRG-05MG, and M059K were obtained from the Bioresource Collection and Research Center (Food Industry Research and Development Institute, Hsinchu, Taiwan).

Techniques: XTT Assay, Control, Staining, Flow Cytometry, Western Blot

Journal: BMC Cancer

Article Title: Valproic acid-induced amphiregulin secretion confers resistance to temozolomide treatment in human glioma cells

doi: 10.1186/s12885-019-5843-6

Figure Lengend Snippet: AR may confer resistance to TMZ in U87MG cells through EGFR activation. a The protein levels of AR in U87MG/shControl, U87MG/shAR#1, and U87MG/shAR#2 cells were examined using Western blot. b Cells were exposed to increasing concentrations of TMZ for 48 h, and subsequently cytotoxicity was evaluated by an XTT-based assay. The percentage of cell viability is shown relative to untreated controls. * P < 0.05 compared with shControl group. c The cell lysates were prepared, and equal amounts of total cell lysates were subjected to Western blot with the indicated antibodies (cleaved PARP and cleaved caspase-3). d Cells were pre-treated with rAR (50 ng/ml) for 8 h, and then further co-treated with TMZ (1500 μM) for 48 h. Cell viability was measured by XTT assay. e The effect of AR on the activation of EGFR family receptors was analyzed by a human EGFR phosphorylation antibody array. f Cells were pre-treated with AR antibodies (1 μg) for 2 h followed by TMZ (500 μM) treatment for another 48 h, and subsequently cytotoxicity was evaluated by an XTT-based assay. The percentage of cell viability is shown relative to untreated controls. * P < 0.05 compared with untreated control. # P < 0.05 compared with TMZ-treated group. Data are representative of at least 3 independent experiments performed in triplicate

Article Snippet: The human glioblastoma cell line U87MG, DBTRG-05MG, and M059K were obtained from the Bioresource Collection and Research Center (Food Industry Research and Development Institute, Hsinchu, Taiwan).

Techniques: Activation Assay, Western Blot, XTT Assay, Phospho-proteomics, Ab Array, Control

Journal: BMC Cancer

Article Title: Valproic acid-induced amphiregulin secretion confers resistance to temozolomide treatment in human glioma cells

doi: 10.1186/s12885-019-5843-6

Figure Lengend Snippet: The expression level of AR was correlated with TMZ resistance in GBM cell lines. a-b The expression levels of AR in each GBM cell lines were evaluated by using quantitative real-time PCR and Western blot analysis. c Cells were treated with increasing concentrations of TMZ for 24 h, and then cell viability was determined by XTT assay. d The protein levels of AR in DBTRG-05MG/shControl, DBTRG-05MG/shAR#1, and DBTRG-05MG/shAR#2 cells were examined using Western blot. e Cells were exposed to increasing concentrations of TMZ for 48 h, and subsequently cytotoxicity was evaluated by an XTT-based assay. The percentage of cell viability is shown relative to untreated controls. * P < 0.05 compared with shControl group. f The cleaved PARP and cleaved caspase-3 were analyzed by Western blot. g-h The expression of AR in U87MG-S (TMZ-sensitive) and parental U87MG was evaluated by Western blot analysis. Cells treated with TMZ (1000 μM) for 24 h, and cell viability was measured by XTT assay. * P < 0.05 compared with U87MG cells. i U87MG cells were cultured in serum withdrawal medium for 24 h, and the effect of serum starvation on AR expression was analyzed by Western blot. Data are representative of at least 3 independent experiments performed in triplicate

Article Snippet: The human glioblastoma cell line U87MG, DBTRG-05MG, and M059K were obtained from the Bioresource Collection and Research Center (Food Industry Research and Development Institute, Hsinchu, Taiwan).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, XTT Assay, Cell Culture

Journal: Oncology Reports

Article Title: miR-1908 as a novel prognosis marker of glioma via promoting malignant phenotype and modulating SPRY4/RAF1 axis

doi: 10.3892/or.2017.6003

Figure Lengend Snippet: miR-1908 promotes glioma cell proliferation and invasion. (A and B) The expression of miR-1908 in U251 cell were affected by transfection of miR-1908 mimics or inhibitor. (C) Downregulated miR-1908 level inhibited the proliferation of U251 cells. (D) miR-1908 significantly promoted the proliferation of U251 cells by >50% at all time-points following 72 h of incubation. (E and F) Upregulated miR-1908 enhanced invasion ability of glioma cells. (G and H) miR-1908 mimics enhanced the activity of MMP-2, while miR-1908 inhibitor reduced its activity.

Article Snippet: Human glioma cell line U251 was purchased from the Cell Resource Center of Beijing Xiehe (Beijing, China) and cultivated in an incubator at 37°C with 5% CO 2 .

Techniques: Expressing, Transfection, Incubation, Activity Assay

Journal: Oncology Reports

Article Title: miR-1908 as a novel prognosis marker of glioma via promoting malignant phenotype and modulating SPRY4/RAF1 axis

doi: 10.3892/or.2017.6003

Figure Lengend Snippet: miR-1908 enhances the ability of glioma cell anti-apoptosis via regulating Bcl-2/Bax expression. (A-D) Double staining with Annexin V-FITC and PI was used to assess apoptosis in U251 cells transfected with miR-1908 mimics or inhibitor, and miR-1908 inhibitor markedly increased cell apoptosis rate. (E) Western blot analysis was performed to determine Bcl-2 and Bax protein expression in U251 cells transfected with miR-1908 mimics or inhibitor, and Bax was increased following transfection with miR-1908 inhibitor, but a marked decrease in the expression of Bcl-2. In contrast, transfection of miR-1908 mimics downregulated the expression level of Bax.

Article Snippet: Human glioma cell line U251 was purchased from the Cell Resource Center of Beijing Xiehe (Beijing, China) and cultivated in an incubator at 37°C with 5% CO 2 .

Techniques: Expressing, Double Staining, Transfection, Western Blot

Journal: Oncology Reports

Article Title: miR-1908 as a novel prognosis marker of glioma via promoting malignant phenotype and modulating SPRY4/RAF1 axis

doi: 10.3892/or.2017.6003

Figure Lengend Snippet: miR-1908 upregulates the expression level of RAF1 through targeting SPRY4. (A and B) PPI analysis of interaction genes with SPRY4, and interaction with pro-oncogene RAF1. (C and D) The expression level of RAF1 was upregulated in glioma tissue compared with normal tissue. (E) The expression level of SPRY4 was negatively correlated with RAF1 in glioma patients from TCGA KIRC database. (F and G) miR-1908 mimics were transfected into U251 cells, the relative mRNA level of SPRY4 and RAF1 were detected by qRT-PCR. (H) Upregulated miR-1908 promoted RAF1 expression via decreasing the expression of SPRY4.

Article Snippet: Human glioma cell line U251 was purchased from the Cell Resource Center of Beijing Xiehe (Beijing, China) and cultivated in an incubator at 37°C with 5% CO 2 .

Techniques: Expressing, Transfection, Quantitative RT-PCR

Journal: Cancer Management and Research

Article Title: The Potential Significance of the EMILIN3 Gene in Augmenting the Aggressiveness of Low-Grade Gliomas is Noteworthy

doi: 10.2147/CMAR.S463694

Figure Lengend Snippet: EMILIN3 gene function experiment in LN229. qRT-PCR transfected LN229 human glioma cells efficiently (A) . The CCK-8 experiment and live/dead staining findings demonstrated that overexpression of the EMILIN3 gene may increase LN229 cell line proliferation and vitality ( B and C ). Overexpression of the EMILIN3 gene enhances the migration, invasiveness, and colony formation of the LN229 cell line (D-F ).

Article Snippet: The LN229 and HS-683 human glioma cell line were purchased from iCell Bioscience Inc, and grown in DMEM media (Gibco, USA) with 10% foetal bovine serum (BI, USA) and 1g/mL penicillin/streptomycin (Hyclone, USA) at 37 degrees Celsius and 5% carbon dioxide.

Techniques: Quantitative RT-PCR, Transfection, CCK-8 Assay, Staining, Over Expression, Migration

Journal: Frontiers in Oncology

Article Title: System analysis based on the migration- and invasion-related gene sets identifies the infiltration-related genes of glioma

doi: 10.3389/fonc.2023.1075716

Figure Lengend Snippet: EMP3 silencing attenuates the migration and invasion of glioma cells. (A) The expression of EMP3 in different cell lines in the CCLE datasets. (B–D) Western blot and qPCR display EMP3 express highly in U118 and A172. (E–G) The growth curve and plate cloning experiment display that EMP3 silencing has little effect on the cell proliferation of U118 and A172. (H, I) EMP3 knockdown inhibits the migration and invasion of glioma cells. ( J–O , 20×) qPCR displays EMP3 silencing attenuates the expression of MMP2 and MMP9 expression (ns, p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001).

Article Snippet: The normal human astroglia (NHA), which were purchased from the Chongqing Golden Magpie Technology Development Co., and U251, LN229, A172, U118, and U87 cells, which were purchased from the China Center for Type Culture Collection (Shanghai, China), were cultured in the DMEM medium supplemented with 10% FBS (HyClone, UT, USA) and 1% penicillin–streptomycin (Beyotime Biotechnology, Jiangsu, China).

Techniques: Migration, Expressing, Western Blot, Cloning, Knockdown

Journal: Frontiers in Oncology

Article Title: System analysis based on the migration- and invasion-related gene sets identifies the infiltration-related genes of glioma

doi: 10.3389/fonc.2023.1075716

Figure Lengend Snippet: EMP3 silencing suppressed EMT marker expression. (A) Western blot analysis of U118 transfected with indicated siRNAs targeting EMP3 (siEMP3#1, siEMP3#2, and siEMP3#3) or siNC. (B) Western blot analysis of A172 transfected with indicated siRNAs targeting EMP3 (siEMP3#1, siEMP3#2, and siEMP3#3) or siNC. (p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001).

Article Snippet: The normal human astroglia (NHA), which were purchased from the Chongqing Golden Magpie Technology Development Co., and U251, LN229, A172, U118, and U87 cells, which were purchased from the China Center for Type Culture Collection (Shanghai, China), were cultured in the DMEM medium supplemented with 10% FBS (HyClone, UT, USA) and 1% penicillin–streptomycin (Beyotime Biotechnology, Jiangsu, China).

Techniques: Marker, Expressing, Western Blot, Transfection