Journal: Journal of Cellular and Molecular Medicine

Article Title: Combination of taxol and Bcl-2 siRNA induces apoptosis in human glioblastoma cells and inhibits invasion, angiogenesis and tumour growth

doi: 10.1111/j.1582-4934.2008.00539.x

Figure Lengend Snippet: In vitro Matrigel invasion assay using U138MG and U251MG cells. Treatment (72 hrs) of cells: transfection with a plasmid vector expressing scrambled siRNA (treated control), 100 nM taxol, transfection with a plasmid vector expressing Bcl-2 siRNA and taxol + Bcl-2 siRNA. Invasion assays were carried out in 12-well transwell inserts of polycarbonate filters. After 48 hrs incubation at 37°C in a CO 2 incubator, the membranes were collected and stained with HEMA. The number of cells that migrated to the undersurface of the membrane were examined under a microscope, counted and photographed. (A) The changes in capability of invasion of U138MG cells after the treatments. (B) Quantitative evaluation of invading U138MG cells. (C) The changes in capability of invasion of U251MG cells after the treatments. (D) Quantitative evaluation of invading U251MG cells. The quantitative data are presented as mean ± S.D. of cells from 10 randomly selected microscopic fields from three independent wells (* P < 0.001 when compared with the scrambled siRNA treatment mean values and # P < 0.001 when compared with taxol or Bcl-2 siRNA treatment mean values).

Article Snippet: Human glioblastoma U138MG cell line was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: In Vitro, Invasion Assay, Transfection, Plasmid Preparation, Expressing, Control, Incubation, Staining, Membrane, Microscopy

Journal: BMC Cancer

Article Title: Impact of PKCε downregulation on autophagy in glioblastoma cells

doi: 10.1186/s12885-018-4095-1

Figure Lengend Snippet: PKCε protein levels in HeLaPKCεA/E and glioblastoma cell lines. Protein extracts from HeLaPKCεA/E (−Dox), HeLaPKCεA/E (+Dox), U-118 MG, U-138 MG, T98G, LN18 cell lines were subjected to Western blot analysis using antibodies against PKCε and GAPDH. HeLaPKCεA/E cells with doxycycline-induced expression of PKCε (+Dox) and without doxycycline treatment (−Dox) served as a reference cell lines with low and high expression of PKCε, respectively. The average relative absorbance of three independent experiments is presented in a bar graph

Article Snippet: Two glioblastoma cell lines (U-138 MG and U-118 MG) were transfected with small interfering RNA against PKCε (PKCε siRNA) and non-targeting siRNA (Control-siRNA) (Santa Cruz Biotechnology, CA, USA).

Techniques: Western Blot, Expressing

Journal: BMC Cancer

Article Title: Impact of PKCε downregulation on autophagy in glioblastoma cells

doi: 10.1186/s12885-018-4095-1

Figure Lengend Snippet: Effect of PKCε downregulation on the adhesion of glioblastoma cells. a . U-138 MG and b U-118 MG cells were transfected for 72 h with PKCε siRNA (PKCε siRNA) and Control siRNA and then were seeded in culture plates coated with Matrigel (Corning Life Sciences, NY, USA). The photos represent cell adhesion under the microscope at 100 x magnification field. c Evaluation of FAK expression in U-138 MG and U-118 MG cells with knockdown PKCε. Total protein expression and FAK phosphorylation at Tyr-397/Tyr-576/577 were assessed. GAPDH was used as a loading control and as an internal standard. Representative blots are shown. The densitometric analysis represents means ±SD of three independent experiments. * P < 0.05, statistically significant compared to Control siRNA

Article Snippet: Two glioblastoma cell lines (U-138 MG and U-118 MG) were transfected with small interfering RNA against PKCε (PKCε siRNA) and non-targeting siRNA (Control-siRNA) (Santa Cruz Biotechnology, CA, USA).

Techniques: Transfection, Control, Microscopy, Expressing, Knockdown, Phospho-proteomics

Journal: Cancer Cell International

Article Title: Glioma exosomal microRNA-148a-3p promotes tumor angiogenesis through activating the EGFR/MAPK signaling pathway via inhibiting ERRFI1

doi: 10.1186/s12935-020-01566-4

Figure Lengend Snippet: miR-148a-3p is highly expressed in glioma tissues, cell lines and glioma cells-derived exosomes. a The box plot showing the miR-148a-3p expression determined by RT-qPCR in non-neoplastic brain tissues of control patients (n = 20) and neoplastic brain tissues of glioma patients (n = 45) including anaplastic astrocytoma, anaplastic oligodendroglioma, oligodendroglioma, astrocytoma, and glioblastoma (U6 was used as internal control). b miR-148a-3p expression determined by RT-qPCR in normal HA and glioma cell lines U-138-MG (U-138), U251-MG (U251), and LN229 (U6 was used as internal control). c Western blot analysis of exosome specific markers (CD63, CD81, and TSG101), and ER stress-related protein CANX in the exosomes extracted from normal HA and glioma cell lines U-138-MG (U-138), U251-MG (U251), and LN229. d Transmission electron microscopic identification of exosomes (scale bar = 100 nm). e NanoSight particle size analysis to quantify exosome concentration and average diameter. f Expression of miR-148a-3p was determined by RT-qPCR in the exosomes extracted from normal HA and glioma cell lines U-138-MG, U251-MG, and LN229 (U6 was used as internal control). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 vs. control brain tissues, HAs or HAs-derived exosomes (HA-exo). Data are shown as mean ± SEM. The cell experiments were performed in 3 repeats and each repeat was performed in technical replicate (triplicate). One-way ANOVA was used for data analysis among multiple groups, followed by Tukey's test

Article Snippet: Fig. 1 miR-148a-3p is highly expressed in glioma tissues, cell lines and glioma cells-derived exosomes. a The box plot showing the miR-148a-3p expression determined by RT-qPCR in non-neoplastic brain tissues of control patients (n = 20) and neoplastic brain tissues of glioma patients (n = 45) including anaplastic astrocytoma, anaplastic oligodendroglioma, oligodendroglioma, astrocytoma, and glioblastoma (U6 was used as internal control). b miR-148a-3p expression determined by RT-qPCR in normal HA and glioma cell lines U-138-MG (U-138), U251-MG (U251), and LN229 (U6 was used as internal control). c Western blot analysis of exosome specific markers (CD63, CD81, and TSG101), and ER stress-related protein CANX in the exosomes extracted from normal HA and glioma cell lines U-138-MG (U-138), U251-MG (U251), and LN229. d Transmission electron microscopic identification of exosomes (scale bar = 100 nm). e NanoSight particle size analysis to quantify exosome concentration and average diameter. f Expression of miR-148a-3p was determined by RT-qPCR in the exosomes extracted from normal HA and glioma cell lines U-138-MG, U251-MG, and LN229 (U6 was used as internal control).

Techniques: Derivative Assay, Expressing, Quantitative RT-PCR, Control, Western Blot, Transmission Assay, Particle Size Analysis, Concentration Assay