Journal: Journal of Cancer

Article Title: Preferential Association of Lissencephaly-1 Gene Expression with CD133+ Glioblastoma Cells

doi: 10.7150/jca.17635

Figure Lengend Snippet: Lis1 and CD133 gene expression in neurosphere-like U87 cells. (A) Exposure of U87 glioma cells to stem-conditioned medium (SM) induces phenotypic modifications resulting in neurospheres after five days, in both regular U87 cells (left) and shLis1-U87 cells (right). The expression of Lis1 (B) and CD133 (D) is induced in U87 cells exposed to SM, with the highest level at day 5 of incubation. (C) As expected, silencing Lis1 gene in U87 cells (shLis-U87) inhibits its induction in cells incubated with SM. CD133 induction is almost abrogated in shLis-U87 cells (E) compared with U87 cells (D). Lis1 is highly expressed in CD133+ cells isolated from U87 cell line and primary glioblastoma (HTC1 and HTC2) cell cultures in normal culture medium (CM) or SM (F) . Data show enrichment up to 60 fold in Lis1 expression in CD133 + fractions for cells grown in CM and up to 32 fold in cells incubated in SM. Lis1 expression in CD133+ fraction isolated from U87 cells grown in CM is 32 times higher than that of CD133- fraction (U87 columns); in CD133+ fraction isolated from U87 incubated in SM Lis1 expression is 35 times higher than in CD133 negative cells from the same culture. The negative control is represented by CD133+ cells isolated from shLis-U87 incubated in CM or SM for which Lis1 was not increased as compared with CD133- cells (shLis-U87/CM and SM columns).

Article Snippet: U87 cells were transfected with a mix of three plasmids, each containing specific shRNA for Lis1 using Plasmid Transfection Reagent in Plasmid Transfection Medium (Santa Cruz, CA).

Techniques: Gene Expression, Expressing, Incubation, Isolation, Negative Control

Journal: Journal of Cancer

Article Title: Preferential Association of Lissencephaly-1 Gene Expression with CD133+ Glioblastoma Cells

doi: 10.7150/jca.17635

Figure Lengend Snippet: Proliferation of irradiated U87 and shLis-U87 cells. Cells having Lis1 silenced or not were irradiated with X-ray doses from 5 to 50 Gy. Cells seeded at a density of 1x10 4 cells/well, in quadruplicates or triplicates in E-plates and placed in xCelligence RTCA instrument, were followed-up for 100 hours (A) . Alternatively, irradiated or not irradiated cells were seeded in 24-well plates and the DNA amount per well was determined using Hoechst 33342 (B) . Both methods showed that irradiated U87 cells recovered better their proliferative capacity than shLis-U87 cells.

Article Snippet: U87 cells were transfected with a mix of three plasmids, each containing specific shRNA for Lis1 using Plasmid Transfection Reagent in Plasmid Transfection Medium (Santa Cruz, CA).

Techniques: Irradiation

Journal: Journal of Cancer

Article Title: Preferential Association of Lissencephaly-1 Gene Expression with CD133+ Glioblastoma Cells

doi: 10.7150/jca.17635

Figure Lengend Snippet: Cell adhesion, migration and proliferation of CD133 + cells isolated from U87 and shLis-U87 cells . CD133+ cells were isolated from control U87 and shLis-U87 cells. The purity of the fraction is revealed by higher CD133 expression (assessed by RT-PCR) in CD133+ fraction as compared with CD133- fraction (A) . CD133+ cells isolated from control U87 (blue circles) or from shLis-U87 (red squares) cultures were subjected to functional assays using xCELLigence Real-Time Cell Analysis instruments. The data representing the recorded cell index at different times show that CD133+ cells isolated from shLis-U87 culture present two times lower (B) adherence to the surface, (C) migratory potential and (D) proliferative rate, as compared with the those isolated from control U87 culture.

Article Snippet: U87 cells were transfected with a mix of three plasmids, each containing specific shRNA for Lis1 using Plasmid Transfection Reagent in Plasmid Transfection Medium (Santa Cruz, CA).

Techniques: Migration, Isolation, Control, Expressing, Reverse Transcription Polymerase Chain Reaction, Functional Assay, Cell Analysis

Journal: Oncology Letters

Article Title: Anticancer effect of the oncolytic Newcastle disease virus harboring the PTEN gene on glioblastoma

doi: 10.3892/ol.2024.14752

Figure Lengend Snippet: mRNA expression and protein expression of PTEN in normal astrocyte or GBM cell lines, and inhibition by rNDV-PTEN of cell viability and migration by inducing apoptotic cell death in U87-MG cells. PTEN (A) mRNA and (B) protein expression in normal astrocyte (CCF-STTG1) and GBM cell lines (U87-MG, T98G, Proneural X01 and Mesenchymal 83). CCF-STTG1 cells and U87-MG cells were infected rNDV or rNDV-PTEN 1 MOI for 36 h. (C) Cell viability assay performed using CCF-STTG1 cells and GBM cell lines with rNDV or rNDV-PTEN 1 MOI treatment using a Cell Counting Kit-8 Kit. (D) U87-MG cells were infected with rNDV or rNDV-PTEN and a Transwell assay was performed to assess cell migration. Cells migrated from the upper chamber to the lower chamber were stained with DAPI. Scale bar, 50 µm. (E) Apoptosis (DNA fragmentation) in U87-MG cells measured using TUNEL staining after virus infection. *P<0.05 vs. CCF-STTG1 or CON. PTEN, phosphatase and tensin homolog; GBM, glioblastoma; rNDV, recombinant Newcastle disease virus; MOI, multiplicity of infection; CON, control; n.s., not significant.

Article Snippet: CCF-STTG1, U87-MG, T98G, Mesenchymal 83 and Proneural X01 cells were seeded at 1×10 4 cells/well in 96-well plates (cat. no. 34096; SPL Life Sciences).

Techniques: Expressing, Inhibition, Migration, Infection, Viability Assay, Cell Counting, Transwell Assay, Staining, TUNEL Assay, Virus, Recombinant, Control