Journal: MedComm

Article Title: Discovery of a First‐in‐Class Murine Double Minute 2‐Recruiting Positive Transcription Elongation Factor B PROTAC Degrader With Selective Antitumor Activity

doi: 10.1002/mco2.70723

Figure Lengend Snippet: MDM2‐ and p53‐dependent activities of MDM2‐recruiting P‐TEFb PROTAC degraders. (A) Effects of siRNA‐mediated MDM2 knockdown on cellular sensitivity to dCDK9‐010. Data were presented as mean ± standard deviation (SD); n = 3. (B) Effects of MDM2 knockdown on dCDK9‐010‐mediated CDK9/Cyclin T degradation. TC‐32 cells were transfected with indicated siRNAs for 48 h, followed by treatment with 2 µM dCDK9‐010 for 8 h. (C) MDM2 mRNA levels in wild type versus TP53 ‐knockout U87 cells, with or without compound treatment (2 µM, 8 h). Data were presented as mean ± SD ( n = 3); *** p < 0.001 based on one‐way analysis of variance (ANOVA); n.s., no significance. (D) Immunoblot analysis of P‐TEFb components, p53, and MDM2 in wild‐type and TP53 ‐knockout isogenic U87 cells after compound treatment (2 µM, 8 h). (E) Viability of isogenic U87 cells treated with dCDK9‐010. Data were presented as mean ± SD ( n = 3). (F) Immunoblot analysis of P‐TEFb components, p53, and MDM2 in NCI‐H226 and TC‐32 cells after siRNA‐mediated TP53 silencing and dCDK9‐010 treatment (2 µM, 8 h). (G) Effects of TP53 or MDM4 knockdown by siRNA relative to control on dCDK9‐010 sensitivity in NCI‐H226 and TC‐32 cells. Data were presented as mean ± SD ( n = 3).

Article Snippet: HEK293T (ATCC), TC‐32 (COG Repository, USA), HCT116 (ATCC), A549 (ATCC), HEPG2 (ATCC), HT1080 (ATCC), MDA‐MB‐231 (ATCC), U87 (U‐87MG, ATCC), U251 (U251MG, ATCC), and MKN45 (ATCC) cells lines were maintained in Dulbecco's modified Eagle medium (DMEM; Sigma‐Aldrich, Taufkirchen, Germany).

Techniques: Knockdown, Standard Deviation, Transfection, Knock-Out, Western Blot, Control

Journal: MedComm

Article Title: Discovery of a First‐in‐Class Murine Double Minute 2‐Recruiting Positive Transcription Elongation Factor B PROTAC Degrader With Selective Antitumor Activity

doi: 10.1002/mco2.70723

Figure Lengend Snippet: Selective activities of compounds 12 (dCDK9‐009) and 13 (dCDK9‐010) against TP53 wild‐type cancer cells. (A and B) Examination of dose‐dependent impacts of dCDK9‐009 and dCDK9‐010 in TP53 wild‐type HCT116, HT1080, NCI‐H460, and U87. Cells were treated with increasing concentrations of indicated compounds for 24 h. (C and D) Examination of impacts of dCDK9‐009 and dCDK9‐010 in non‐malignant HEK293T and mesenchymal stem cell line ASC52telo. Cells were treated with increasing concentrations of indicated compounds for 24 h. (E and F) Effect of dCDK9‐009 and dCDK9‐010 treatment on cellular viabilities of indicated cancer cell lines, as well as ASC52telo. IC 50 was presented as mean ± SD with three independent replicates. (G) Heatmap summarizing the IC 50 values of dCDK9‐009 and dCDK9‐010 across human cell lines tested in this study (related to E and F, Figures S3, S4, and S8).

Article Snippet: HEK293T (ATCC), TC‐32 (COG Repository, USA), HCT116 (ATCC), A549 (ATCC), HEPG2 (ATCC), HT1080 (ATCC), MDA‐MB‐231 (ATCC), U87 (U‐87MG, ATCC), U251 (U251MG, ATCC), and MKN45 (ATCC) cells lines were maintained in Dulbecco's modified Eagle medium (DMEM; Sigma‐Aldrich, Taufkirchen, Germany).

Techniques:

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: Cancer Cell International

Article Title: Bifidobacterium enhances the antitumor efficacy of carboplatin in glioblastoma cells: targeting apoptotic and cell cycle regulatory pathways via Caspase , AKT/PTEN , and P53/P21 signaling

doi: 10.1186/s12935-025-04099-w

Figure Lengend Snippet: Evaluation of U-87 MG Cell Viability Following Treatment with Bifidobacterium reuteri Ab.338 SH (B.R). Glioblastoma U-87 MG cells were treated with B.R at OD600 values of 0.8, 1.0, 1.25, and 1.5 (10 µL/mL). After 24 h of exposure, cell viability was assessed using the MTT assay. Results are presented as mean ± SD from three independent biological replicates. Differences compared to untreated controls were analyzed for statistical significance (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The human glioblastoma cell line U-87 MG and human umbilical vein endothelial cells (HUVECs) were obtained from the Pasteur Institute of Iran (National Cell Bank).

Techniques: MTT Assay

Journal: Cancer Cell International

Article Title: Bifidobacterium enhances the antitumor efficacy of carboplatin in glioblastoma cells: targeting apoptotic and cell cycle regulatory pathways via Caspase , AKT/PTEN , and P53/P21 signaling

doi: 10.1186/s12935-025-04099-w

Figure Lengend Snippet: Cytotoxic Effects of Carboplatin (C.P) on U-87 MG Cells. U-87 MG cells were treated with increasing concentrations of C.P (16.5, 18.5, 20.5, and 22.5 µg/mL) for 24 h. Viability was determined using the MTT assay. The data, derived from three independent experiments ( n = 3), are reported as mean ± SD. Statistically significant differences compared to untreated cells are indicated (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The human glioblastoma cell line U-87 MG and human umbilical vein endothelial cells (HUVECs) were obtained from the Pasteur Institute of Iran (National Cell Bank).

Techniques: MTT Assay, Derivative Assay

Journal: Cancer Cell International

Article Title: Bifidobacterium enhances the antitumor efficacy of carboplatin in glioblastoma cells: targeting apoptotic and cell cycle regulatory pathways via Caspase , AKT/PTEN , and P53/P21 signaling

doi: 10.1186/s12935-025-04099-w

Figure Lengend Snippet: Synergistic Cytotoxicity of Carboplatin and Bifidobacterium reuteri Ab.338 SH in U-87 MG Cells. Cells were co-treated with fixed B.R (OD600 = 1.0) and varying concentrations of C.P (128, 138, 148, and 158 ng/mL). Following 24-hour incubation, cell viability was assessed via MTT assay. Data are presented as mean ± SD from three separate experiments. Significant differences relative to the control group are denoted (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The human glioblastoma cell line U-87 MG and human umbilical vein endothelial cells (HUVECs) were obtained from the Pasteur Institute of Iran (National Cell Bank).

Techniques: Incubation, MTT Assay, Control

Journal: Cancer Cell International

Article Title: Bifidobacterium enhances the antitumor efficacy of carboplatin in glioblastoma cells: targeting apoptotic and cell cycle regulatory pathways via Caspase , AKT/PTEN , and P53/P21 signaling

doi: 10.1186/s12935-025-04099-w

Figure Lengend Snippet: Apoptosis-Related Gene Expression in U-87 MG Cells Treated with C.P + B.R at IC₅₀. Quantitative real-time PCR was performed on cells treated with the IC₅₀ dose of C.P (148 ng/mL) in combination with B.R (OD600 = 1.0) for 24 h. The analysis revealed significant upregulation of pro-apoptotic genes (Caspase-3, −8, −9, Bax, PTEN, P53, P21, IκB, and Fas), while anti-apoptotic Bcl-2 and components of the PI3K/AKT/mTOR pathway (AKT and mTOR) were significantly downregulated. Results are shown as mean ± SD from three biological replicates. Statistical comparisons to control: * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The human glioblastoma cell line U-87 MG and human umbilical vein endothelial cells (HUVECs) were obtained from the Pasteur Institute of Iran (National Cell Bank).

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Control

Journal: Cancer Cell International

Article Title: Bifidobacterium enhances the antitumor efficacy of carboplatin in glioblastoma cells: targeting apoptotic and cell cycle regulatory pathways via Caspase , AKT/PTEN , and P53/P21 signaling

doi: 10.1186/s12935-025-04099-w

Figure Lengend Snippet: Schematic representation of the molecular mechanism underlying the synergistic apoptotic effects of Bifidobacterium reuteri Ab.338 SH and Carboplatin in U-87 MG cells. The combined treatment enhances apoptotic signaling through multiple pathways. Activation of Fas and upregulation of PTEN lead to Caspase-8 and Caspase-9 activation via the extrinsic pathway. Simultaneously, downregulation of AKT signaling promotes intrinsic apoptosis through sequential activation of Caspase-3 and Caspase-9 , ultimately leading to BAX activation and cell death. The co-treatment also suppresses AKT -mediated cell survival signaling and inhibits the expression of anti-apoptotic BCL-2 via P53/P21 axis modulation. Together, these molecular events contribute to enhanced apoptosis and reduced survival in glioblastoma cells. ↑ indicates upregulation; ↓ indicates downregulation

Article Snippet: The human glioblastoma cell line U-87 MG and human umbilical vein endothelial cells (HUVECs) were obtained from the Pasteur Institute of Iran (National Cell Bank).

Techniques: Activation Assay, Expressing

Journal: Cancer Cell International

Article Title: Bifidobacterium enhances the antitumor efficacy of carboplatin in glioblastoma cells: targeting apoptotic and cell cycle regulatory pathways via Caspase , AKT/PTEN , and P53/P21 signaling

doi: 10.1186/s12935-025-04099-w

Figure Lengend Snippet: Flow Cytometric Detection of Apoptosis in U-87 MG Cells Treated with C.P + B.R at IC₅₀ Concentration. U-87 MG cells were exposed to a combination of C.P (148 ng/mL) and B.R (OD600 = 1.0) for 24 h, followed by Annexin V-FITC/PI staining and flow cytometry. Representative dot plots display four distinct populations: necrotic (Q1: Annexin⁻/PI⁺), late apoptotic (Q2: Annexin⁺/PI⁺), early apoptotic (Q3: Annexin⁺/PI⁻), and viable (Q4: Annexin⁻/PI⁻). Combined treatment markedly increased both early and late apoptotic populations compared to untreated controls. Each plot is representative of three independent experiments

Article Snippet: The human glioblastoma cell line U-87 MG and human umbilical vein endothelial cells (HUVECs) were obtained from the Pasteur Institute of Iran (National Cell Bank).

Techniques: Concentration Assay, Staining, Flow Cytometry

Journal: Cancer Cell International

Article Title: Bifidobacterium enhances the antitumor efficacy of carboplatin in glioblastoma cells: targeting apoptotic and cell cycle regulatory pathways via Caspase , AKT/PTEN , and P53/P21 signaling

doi: 10.1186/s12935-025-04099-w

Figure Lengend Snippet: Quantification of Cell Death Stages in U-87 MG Cells Following Combined C.P + B.R Treatment. Bar graph displays the distribution of viable, early apoptotic, late apoptotic, and necrotic U-87 MG cells after 24-hour exposure to the IC₅₀ dose of C.P (148 ng/mL) plus B.R (OD600 = 1.0), as determined by flow cytometry. Early apoptosis increased from 0.0% (control) to 27.64%, and late apoptosis rose to 39.78%, while necrosis remained minimal. Data are expressed as mean ± SEM from three biological replicates, with statistical significance indicated (* p < 0.05), confirming apoptosis as the predominant mechanism of cytotoxicity

Article Snippet: The human glioblastoma cell line U-87 MG and human umbilical vein endothelial cells (HUVECs) were obtained from the Pasteur Institute of Iran (National Cell Bank).

Techniques: Flow Cytometry, Control