Journal: The Journal of Biological Chemistry

Article Title: Transcriptome analysis of polyploid giant cancer cells and their progeny reveals a functional role for p21 in polyploidization and depolyploidization

doi: 10.1016/j.jbc.2024.107136

Figure Lengend Snippet: Expression of p21 in PGCC. Western blot analysis was performed to verify the upregulation of p21 mRNA at the protein level in PGCC compared to untreated cancer cells (UT) ( A and B ). Three independently performed experiments were analyzed for expression of p21 relative to GAPDH. ∗ p < 0.05, ∗∗ p < 0.005. ( C – E ) Distribution of p21 in untreated cells and PGCC derived from PPC1, U118MG, and HT29 cells. Similar results were obtained in three independent experiments.

Article Snippet: U118MG and HT29 cells were obtained from Mediatech and ATCC, respectively.

Techniques: Expressing, Western Blot, Derivative Assay

Journal: The Journal of Biological Chemistry

Article Title: Transcriptome analysis of polyploid giant cancer cells and their progeny reveals a functional role for p21 in polyploidization and depolyploidization

doi: 10.1016/j.jbc.2024.107136

Figure Lengend Snippet: The effect of UC2288 on depolyploidization and polyploidization. A and B , colony formation assay of PGCC derived from PPC1 ( A ) or U118MG cells ( B ). Data are from three independent experiments with 2 to 4 technical replicates. C and D , analysis of DNA content following a 3-days stress exposure. C , representative flow cytometry traces. D , quantification of flow cytometry data from three independent experiments. Two-way Anova indicated differences between groups and was followed up with a unpaired, 2-tailed t test to evaluate differences between groups; ∗ p < 0.05; ∗∗ p < 0.005, ∗∗∗ p < 0.0005.

Article Snippet: U118MG and HT29 cells were obtained from Mediatech and ATCC, respectively.

Techniques: Colony Assay, Derivative Assay, Flow Cytometry

Journal: British Journal of Cancer

Article Title: H1/pHGFK1 nanoparticles exert anti-tumoural and radiosensitising effects by inhibition of MET in glioblastoma

doi: 10.1038/bjc.2017.461

Figure Lengend Snippet: The anti-tumoural and radiosensitising functions of H1/pHGFK1 nanoparticles in the orthotopic luciferase-labelling U118 cells-xenografted nude mouse model. ( A , C ) Representative bioluminescent images of tumour-bearing live mice treated with PBS-, H1/pEGFP, IR, IR+ H1/pEGFP, H1/pHGFK1 and IR+ H1/pHGFK1 were captured by in vivo imaging on day 7 and day 14 post-treatment, respectively. Red circles marking the region of interests (ROIs) in each mouse; bioluminescent intensities (BIs, CPS: counts per second) are in the red boxes. ( B , D ) Statistical histograms of ROIs in living mice from the six groups on days 7 and 14 post-treatment, respectively (Student’s t -test, n =6, * P <0.05; ** P <0.01; *** P <0.001). ( E ) The survival curve of the tumour-bearing mice in the six groups (Student’s t -test, n =6, ** P <0.01).

Article Snippet: This luciferase-stable U118 cell line was established using Lenti-luciferase (Cat. D13GZ; GenePharma, Shanghai, China) according to the manufacturer’s instructions.

Techniques: Luciferase, In Vivo Imaging

Journal: BMC Cancer

Article Title: A novel chalcone derivative which acts as a microtubule depolymerising agent and an inhibitor of P-gp and BCRP in in-vitro and in-vivo glioblastoma models

doi: 10.1186/1471-2407-9-242

Figure Lengend Snippet: Effects of JAI-51 on a panel of glioblastoma cell lines . U118, U138, U373, LN229 cells were cultured for 24 h before the addition of JAI-51 (1 to 10 μM). Cells were used for either cell proliferation tests using MTT at the indicated times (A) or for cell cycle analysis at 24 h (B) . (A) After 24 h induction time with 10 μM JAI-51, the decrease in proliferation was statistically significant for all the cell lines tested compared to the untreated cells ( P < 0.05 treated versus control), and after 48 h induction time, this decrease in proliferation was also significant with 5 μM JAI-51 for all the cell lines tested ( P < 0.05 treated versus control). The results are expressed as the mean ± SD from 3 different experiments. Mitotic index (M.I.) (C, left panel) was determined at 24 h on slide after Papanicolaou staining, and apoptosis (C, right panel) was determined at 48 h using the Annexin V-FITC/PI method (* P < 0.05 treated versus control). (D) Photomicrographs of control LN229 and GL26 cells and cells treated with 10 μM JAI-51 for 24 h (LN229+JAI-51, GL26+JAI-51). Treated LN229 and GL26 cells displayed polylobated nuclei and most of the mitotic figures showed numerous abnormalities (complex spindle with several poles). (Magnification × 600).

Article Snippet: Human glioblastoma derived cell lines U118, U138, U373 and LN229 were generously provided by Pr F. Berger (INSERM U318).

Techniques: Cell Culture, Cell Cycle Assay, Control, Staining

Journal: BMC Cancer

Article Title: A novel chalcone derivative which acts as a microtubule depolymerising agent and an inhibitor of P-gp and BCRP in in-vitro and in-vivo glioblastoma models

doi: 10.1186/1471-2407-9-242

Figure Lengend Snippet: Effect of JAI-51 on the apoptotic process in the glioblastoma cell lines.

Article Snippet: Human glioblastoma derived cell lines U118, U138, U373 and LN229 were generously provided by Pr F. Berger (INSERM U318).

Techniques: Activity Assay

Journal: Neuro-Oncology

Article Title: The novel chromatin architectural regulator SND1 promotes glioma proliferation and invasion and predicts the prognosis of patients

doi: 10.1093/neuonc/noz038

Figure Lengend Snippet: SND1 facilitates GBM cell invasion and proliferation. (A) Western blotting results of the SND1 level in U118MG and primary GBM cells of control (scramble/vector), SND1-knockdown (SND1-sh1/vector, SND1-sh2/vector), and SND1 rescue expression (SND1-sh1/SND1, SND1-sh2/SND1) groups. (B) The transwell results of aforementioned U118MG and primary GBM cells. (C) MTT proliferation assays of aforementioned cells. (D) Colony formation of U118MG cells expressing control shRNA (scramble) or SND1-shRNAs (SND1-sh1, sh2). *P < 0.05, **P < 0.01. (E) U118MG and primary GBM cells bearing control shRNA (scramble) or shRNAs against SND1 (SND1-sh1, sh2) were inoculated orthotopically into NOD-SCID mice (n = 5). Tumor volumes were measured by the ISVS system after 28 days (U118MG) or 21 days (primary GBM) of initial implantation. (F) IHC staining for MKI67 (upper panel) in a xenograft tumor from mice transplanted with U118MG and primary GBM cells bearing control shRNA (scramble) or shRNAs against SND1 (SND1-sh1, sh2). Representative images of the invasion condition in HE stained tissue from the scramble, SND1-sh1, and SND1-sh2 (middle and bottom panel) xenografts. The red dashed line marks the tumor border, and the red arrow displays the invasion distance. (G) Survival results analyzed by the KM estimator showed that mice bearing SND1-sh1 and sh2 GBM cells survived longer than the control mice (scramble, P < 0.001).

Article Snippet: The U118MG and primary GBM cells expressing luciferase were implanted (7.5 × 10 4 cells) into the brains of 6-week-old non-obese diabetic, severe combined immunodeficiency (NOD-SCID) mice (Charles River Laboratories).

Techniques: Western Blot, Control, Plasmid Preparation, Knockdown, Expressing, shRNA, Immunohistochemistry, Staining

Journal: Neuro-Oncology

Article Title: The novel chromatin architectural regulator SND1 promotes glioma proliferation and invasion and predicts the prognosis of patients

doi: 10.1093/neuonc/noz038

Figure Lengend Snippet: SND1 knockdown represses a cluster of cell proliferation and motility genes. (A) Functional profiling of differentially expressed genes among U118MG and primary GBM cells with scrambled shRNAs (scramble) or SND1 shRNAs (SND1-sh1 and sh2) transfection (n = 3 for each group). The 2 arrowheads label 32 upregulated genes and 49 downregulated genes after SND1 knockdown (SND1-sh1, sh2). Representative SND1-sh promoted (red) and SND1-sh inhibited genes (green) are shown (left) based on their function (right). (B) The 5 most important pathways enriched among the SND1 candidate regulatory targets are shown in the red box. See also Supplementary Table 10. (C) Venn diagram shows SND1-activated downstream target genes. (D) Network plot illustrating relationships among SND1, RhoA (bold red circle), and downstream pathway members (bold black circle).

Article Snippet: The U118MG and primary GBM cells expressing luciferase were implanted (7.5 × 10 4 cells) into the brains of 6-week-old non-obese diabetic, severe combined immunodeficiency (NOD-SCID) mice (Charles River Laboratories).

Techniques: Knockdown, Functional Assay, Transfection

Journal: Neuro-Oncology

Article Title: The novel chromatin architectural regulator SND1 promotes glioma proliferation and invasion and predicts the prognosis of patients

doi: 10.1093/neuonc/noz038

Figure Lengend Snippet: SND1 upregulates RhoA transcription by inducing long-range chromatin conformation remodeling on the RhoA promoter. (A) SND1-binding sites predicted in the RhoA promoter (top figure) and ChIP-qPCR analyses of the capacities for them to bind with SND1. (B) Binding between SND1 and the RhoA promoter verified by EMSA (Lanes 2, 4 and 5). (C and D) 3C results showed the cross-linking efficiency between SND1 binding sites R1 and R2 in both U118MG and primary GBM cells with or without SND1 knockdown (R1 as the anchor, C; R2 as the anchor, D). DpnII digestion sites are labeled by the vertical lines, and the arrows represent the individual PCR primers. **P < 0.01. (E) Schematic displaying the assembling process of the transcription activation complex on the RhoA promoter.

Article Snippet: The U118MG and primary GBM cells expressing luciferase were implanted (7.5 × 10 4 cells) into the brains of 6-week-old non-obese diabetic, severe combined immunodeficiency (NOD-SCID) mice (Charles River Laboratories).

Techniques: Binding Assay, ChIP-qPCR, Knockdown, Labeling, Activation Assay

Journal: Neuro-Oncology

Article Title: The novel chromatin architectural regulator SND1 promotes glioma proliferation and invasion and predicts the prognosis of patients

doi: 10.1093/neuonc/noz038

Figure Lengend Snippet: SND1 accelerates the GBM cell cycle, proliferation, and invasion in vitro via upregulating RhoA. (A) Western blot results of SND1, RhoA, CCND1, CCNE1, CDK4, and CDKN1B in U118MG and primary GBM cells of the control (scramble) and SND1-knockdown (SND1-sh1, sh2) groups. (B) Cell cycle analyses by flow cytometry of the cells described above. (C) Quantitative RT-PCR detection of SND1 and RhoA mRNA levels in U118MG and primary GBM cells of the scramble/vector, SND1-sh1/vector, SND1-sh2/vector, scramble/RhoA, SND1-sh1/ RhoA, and SND1-sh2/ RhoA groups. (D) Western blot results of SND1, RhoA, CCND1, CCNE1, CDK4, and CDKN1B in the cells described in (C). (E) Results of transwell invasive cell counts of the cells as indicated in (C). (F) MTT proliferation assay of the cells described above. *P < 0.05, **P < 0.01.

Article Snippet: The U118MG and primary GBM cells expressing luciferase were implanted (7.5 × 10 4 cells) into the brains of 6-week-old non-obese diabetic, severe combined immunodeficiency (NOD-SCID) mice (Charles River Laboratories).

Techniques: In Vitro, Western Blot, Control, Knockdown, Flow Cytometry, Quantitative RT-PCR, Plasmid Preparation, Proliferation Assay

Journal: Neuro-Oncology

Article Title: The novel chromatin architectural regulator SND1 promotes glioma proliferation and invasion and predicts the prognosis of patients

doi: 10.1093/neuonc/noz038

Figure Lengend Snippet: SND1 accelerates GBM in vivo growth via RhoA; the RhoA levels indicate high glioma grades and poor prognoses. (A) U118MG and primary GBM cells with empty vector plus scramble control shRNA (scramble/vector) or SND1 shRNA (SND1-sh1/vector, SND1-sh2/vector), or ectopic RhoA plus scrambled shRNA (scramble/RhoA) or SND1 shRNA (SND1-sh1/RhoA, SND1-sh2/RhoA) were inoculated orthotopically into NOD-SCID mice (n = 5). (B) Tumors were measured by the IVIS imaging system. (C) Survival analysis results of the tumor transplant experiment. (D) Representative HE-stained images of the invasion condition among the cells described in (A). Scale bar, 100 μm. (E) IHC results of RhoA expression. Scale bar, 50 μm. (F) Comparing the RhoA LIs among different WHO grades in 187 gliomas and 20 normal brain tissue samples. (G) Linear regression results showed the correlation between SND1 and RhoA in 187 gliomas. (H) Survival analysis results by RhoA LI in WHO grades II–IV glioma samples (left) and grade IV GBM samples (right).

Article Snippet: The U118MG and primary GBM cells expressing luciferase were implanted (7.5 × 10 4 cells) into the brains of 6-week-old non-obese diabetic, severe combined immunodeficiency (NOD-SCID) mice (Charles River Laboratories).

Techniques: In Vivo, Plasmid Preparation, Control, shRNA, Imaging, Staining, Expressing