Journal: Cancer Genomics & Proteomics

Article Title: γ-Glutamylcyclotransferase Depletion Induces p15INK4b and p21Cip1-mediated Senescence via TGF-β2/SMAD3 Pathway Activation in Breast Cancer Cells

doi: 10.21873/cgp.20571

Figure Lengend Snippet: Knockdown of GGCT inhibited the proliferation and caused significant changes in gene expression profiles detected by transcriptomic highthroughput sequencing in MCF-7 cells. (A) mRNA expression of GGCT was analyzed by qRT-PCR 3 days after transfection. n=3 per group; *p<0.05, **p<0.01, and ***p<0.001 using two-tailed Student’s t-test for pairwise comparisons. (B) Western blotting analysis of GGCT and α-tubulin 4 days after transfection of MCF-7 cells with GGCT-siRNA or non-target control siRNA. (C) The relative survival number of trypan blue-negative viable MCF-7 cells at 1, 4, 7 days post-transfection. n=3 per group; *p<0.05, **p<0.01, and ***p<0.001 using one-way ANOVA followed by Tukey’s post hoc test. (D) Representative images at 4 days post-transfection. (E) Hierarchical clustering analysis of differentially expressed genes (DEGs) detected by RNA-seq analysis. The color scale indicates log10(FPKM) and intensity increases from green to red, indicating down-regulation and up-regulation, respectively. (F) Numbers of significantly up-regulated (red) and down-regulated (green) DEGs in siGGCT-transfected MCF-7 cells at days 1, 2, and 3 post-transfection, identified by RNA-seq analysis using the filtering criteria of |log₂ fold change| >1 and q-value <0.01. Scale bar: 50 μm. ANOVA, Analysis of variance; FC, fold change; FPKM, fragments per kilobase of exon model per million mapped reads; GGCT, γ-glutamylcyclotransferase; qRT-PCR, quantitative reversetranscription- polymerase chain reaction; siRNA, small-interfering RNA.

Article Snippet: The breast cancer cell line MCF-7 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (D-MEM; FUJIFILM Wako, Osaka, Japan) supplemented with 10% fetal bovine serum (HyClone, South Logan, UT, USA) and 1% penicillin and streptomycin.

Techniques: Knockdown, Gene Expression, High Throughput Screening Assay, Sequencing, Expressing, Quantitative RT-PCR, Transfection, Two Tailed Test, Western Blot, Control, RNA Sequencing, Polymerase Chain Reaction, Small Interfering RNA

Journal: Cancer Genomics & Proteomics

Article Title: γ-Glutamylcyclotransferase Depletion Induces p15INK4b and p21Cip1-mediated Senescence via TGF-β2/SMAD3 Pathway Activation in Breast Cancer Cells

doi: 10.21873/cgp.20571

Figure Lengend Snippet: GO and KEGG pathway enrichment analyses of DEGs in MCF-7 cells after GGCT knockdown. (A) GO enrichment analysis for biological processes (up-regulated and down-regulated genes), and KEGG pathway analysis of DEGs, using samples collected 3 days after knockdown. The top 10 biological process terms ranked according to p-Values are shown. (B) Hypothetical schema of cell cycle regulation following GGCT knockdown in MCF-7 cells, constructed based on DEGs enriched in the KEGG pathway ‘hsa04110: Cell cycle’ (http://www.kegg.jp). (C) Relative mRNA expression patterns of GGCT, TGF-β2, CDKN1A (p21 Cip1 ), and CDKN2B (p15 INK4b ) were measured with qRT-PCR. n=3 per group; *p<0.05, **p<0.01, and ***p<0.001 using two-tailed Student’s t-test for pairwise comparisons. CDKN1A, Cyclin dependent kinase inhibitor 1A; CDKN2B, cyclin dependent kinase inhibitor 2B; DEGs, differentially expressed genes; GGCT, γ-glutamylcyclotransferase; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; qRT-PCR, quantitative reverse-transcription-polymerase chain reaction; TGF-β2, transforming growth factor-β2.

Article Snippet: The breast cancer cell line MCF-7 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (D-MEM; FUJIFILM Wako, Osaka, Japan) supplemented with 10% fetal bovine serum (HyClone, South Logan, UT, USA) and 1% penicillin and streptomycin.

Techniques: Knockdown, Construct, Expressing, Quantitative RT-PCR, Two Tailed Test, Reverse Transcription, Polymerase Chain Reaction

Journal: Cancer Genomics & Proteomics

Article Title: γ-Glutamylcyclotransferase Depletion Induces p15INK4b and p21Cip1-mediated Senescence via TGF-β2/SMAD3 Pathway Activation in Breast Cancer Cells

doi: 10.21873/cgp.20571

Figure Lengend Snippet: Up-regulation of p15 INK4b and p21 Cip1 contributes to G0/G1 cell cycle arrest and subsequent cell growth inhibition in GGCT-depleted MCF-7 cells. (A) The levels of p15I NK4b and p21 Cip1 proteins were normalized to levels of α-tubulin and the fold change relative to the control was calculated at 4 days after the indicated siRNA transfection. (B) The cell cycle distribution and (C) representative histograms in FACS analysis in MCF-7 cells 4 days after transfection with the indicated siRNAs. n=3 per group; *p<0.05; **p<0.01, and ***p<0.001 vs. control, †p<0.05; ††p<0.01, and †††p<0.001 vs. GGCT, one-way ANOVA followed by Tukey’s post hoc test. (D) The number of viable cells in the trypan blue dye exclusion test and (E) representative images of MCF-7 cells at 7 days after transfection with the indicated siRNAs. Scale bar: 50 μm; n=3 per group; *p<0.05, **p<0.01, and ***p<0.001, using one-way ANOVA followed by Tukey’s post hoc test. ANOVA, Analysis of variance; FACS, fluorescence-activated cell sorting; GGCT, γ-glutamylcyclotransferase; PCR, polymerase chain reaction; siRNA, small-interfering RNA.

Article Snippet: The breast cancer cell line MCF-7 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (D-MEM; FUJIFILM Wako, Osaka, Japan) supplemented with 10% fetal bovine serum (HyClone, South Logan, UT, USA) and 1% penicillin and streptomycin.

Techniques: Inhibition, Control, Transfection, Fluorescence, FACS, Polymerase Chain Reaction, Small Interfering RNA

Journal: Cancer Genomics & Proteomics

Article Title: γ-Glutamylcyclotransferase Depletion Induces p15INK4b and p21Cip1-mediated Senescence via TGF-β2/SMAD3 Pathway Activation in Breast Cancer Cells

doi: 10.21873/cgp.20571

Figure Lengend Snippet: TGF-β2/SMAD signaling axis regulates p15 INK4b and p21 Cip1 expression in GGCT-depleted MCF-7 cells. (A) mRNA expression levels of GGCT and TGF-β2 were analyzed by qRT-PCR 3 days after transfection with GGCT- and/or TGF-β2-siRNA. (B) Western blot analysis of GGCT and TGF-β2 expression 4 days after transfection with GGCT- and/or TGF-β2-siRNA. (C) Western blotting analysis of p15 INK4b , p21 Cip1 , phospho-SMAD2 (pSMAD2), SMAD2, phospho-SMAD3 (pSMAD3), SMAD3, GGCT, and α-tubulin in MCF-7 at 4 days after transfection with GGCT-siRNA and/or TGF-β2-siRNA, or non-target control siRNA. (D) Western blotting analysis of p15 INK4b , p21 Cip1 , pSMAD3, SMAD3, GGCT, and α-tubulin in MCF-7 cells at 4 days after transfection with GGCT-siRNA and/ or SMAD3-siRNA, or non-target control siRNA. (E) The number of viable cells in the trypan blue dye exclusion test and (F) representative images of MCF- 7 cells at 4 days after transfection with the indicated siRNAs. Scale bar: 200 μm; n=3 per group; *p<0.05, **p<0.01, and ***p<0.001 vs. control, †p<0.05; ††p<0.01, and †††p<0.001 vs. GGCT, one-way ANOVA followed by Tukey’s post hoc test. ANOVA, Analysis of variance; GGCT, γ-glutamylcyclotransferase; pSMAD, phosphorylated SMAD; qRT-PCR, quantitative reverse-transcription-polymerase chain reaction; siRNA, small-interfering RNA.

Article Snippet: The breast cancer cell line MCF-7 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (D-MEM; FUJIFILM Wako, Osaka, Japan) supplemented with 10% fetal bovine serum (HyClone, South Logan, UT, USA) and 1% penicillin and streptomycin.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Control, Reverse Transcription, Polymerase Chain Reaction, Small Interfering RNA

Journal: Cancer Genomics & Proteomics

Article Title: γ-Glutamylcyclotransferase Depletion Induces p15INK4b and p21Cip1-mediated Senescence via TGF-β2/SMAD3 Pathway Activation in Breast Cancer Cells

doi: 10.21873/cgp.20571

Figure Lengend Snippet: p15 INK4b , p21 Cip1 , and their upstream regulator TGF-β2 are involved in the induction of cellular senescence following GGCT-KD in MCF-7 cells. (A) Representative images of SA-β-Gal staining 4 days after transfection with the indicated siRNAs, including simultaneous knockdown of GGCT, p15, and p21. Scale bar: 50 μm. (B) The proportion of SA-β-Gal-positive cells in MCF-7 cells are shown. (C) Representative images of SA-β-Gal staining at 4 days after transfection with the indicated siRNAs, including simultaneous knockdown of GGCT and TGF-β2. Scale bar: 50 μm. (D) The proportion of SA-β-Gal-positive cells in MCF-7 cells are shown. n=3 per group; *p<0.05, **p<0.01, ***p<0.001, using one-way ANOVA followed by Tukey’s post hoc test. ANOVA, Analysis of variance; GGCT, γ-glutamylcyclotransferase; KD, knockdown; SA-β-Gal, senescence-associated β-galactosidase; siRNA, smallinterfering RNA; TGF-β2, transforming growth factor-β2.

Article Snippet: The breast cancer cell line MCF-7 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (D-MEM; FUJIFILM Wako, Osaka, Japan) supplemented with 10% fetal bovine serum (HyClone, South Logan, UT, USA) and 1% penicillin and streptomycin.

Techniques: Staining, Transfection, Knockdown

Journal: AMB Express

Article Title: Bioactive potential of Pulicaria undulata from arid regions against multidrug-resistant pathogens and cancer cells

doi: 10.1186/s13568-026-02020-w

Figure Lengend Snippet: Effect of P. undulata extract onto Vero ATCC CCL- normal cells and MCF-7 ATCC HTB-22 breast cancer cell lines, A Cytotoxicity; B Cell viability. Data are presented as Mean ± SE ( n = 3)

Article Snippet: The P. undulata extract cytotoxicity was tested against a panel of Vero ATCC CCL-81cercopithecus aethiops kidney normal cells and MCF-7 ATCC HTB-22 Homo sapiens human breast cancer cell lines, with the findings shown in Fig. , respectively.

Techniques:

Journal: AMB Express

Article Title: Bioactive potential of Pulicaria undulata from arid regions against multidrug-resistant pathogens and cancer cells

doi: 10.1186/s13568-026-02020-w

Figure Lengend Snippet: P. undulata extract effect onto Vero cells and MCF-7 cancer cells morphological features for 48 h (bar scale 10 nm)

Article Snippet: The P. undulata extract cytotoxicity was tested against a panel of Vero ATCC CCL-81cercopithecus aethiops kidney normal cells and MCF-7 ATCC HTB-22 Homo sapiens human breast cancer cell lines, with the findings shown in Fig. , respectively.

Techniques:

Journal: AMB Express

Article Title: Bioactive potential of Pulicaria undulata from arid regions against multidrug-resistant pathogens and cancer cells

doi: 10.1186/s13568-026-02020-w

Figure Lengend Snippet: Effect of P. undulata extract onto Vero ATCC CCL- normal cells and MCF-7 ATCC HTB-22 breast cancer cell lines, A Cytotoxicity; B Cell viability. Data are presented as Mean ± SE ( n = 3)

Article Snippet: Fig. 5 Effect of P. undulata extract onto Vero ATCC CCL- normal cells and MCF-7 ATCC HTB-22 breast cancer cell lines, A Cytotoxicity; B Cell viability.

Techniques:

Journal: AMB Express

Article Title: Bioactive potential of Pulicaria undulata from arid regions against multidrug-resistant pathogens and cancer cells

doi: 10.1186/s13568-026-02020-w

Figure Lengend Snippet: P. undulata extract effect onto Vero cells and MCF-7 cancer cells morphological features for 48 h (bar scale 10 nm)

Article Snippet: Fig. 5 Effect of P. undulata extract onto Vero ATCC CCL- normal cells and MCF-7 ATCC HTB-22 breast cancer cell lines, A Cytotoxicity; B Cell viability.

Techniques:

Journal: RSC Advances

Article Title: Efficient removal of the ciprofloxacin drug using an electrospun Co/Al-layered double hydroxide-embedded chitosan/polyvinylidene fluoride nanofiber membrane

doi: 10.1039/d5ra08795c

Figure Lengend Snippet: Effectiveness of the CS/PVDF and Co/Al-LDH CS/PVDF nanofiber membranes regarding (a) HePG-2 cell viability, (b) HePG-2 inhibitory percentage, (c) MCF-7 cell viability, and (d) MCF-7 inhibitory percentage.

Article Snippet: More specifically, the MCF-7 cell line, which stands for human breast adenocarcinoma, and HepG-2, representing human hepatocellular carcinoma, were both supplied by the American Type Culture Collection (ATCC located at Manassas Virginia USA).

Techniques:

Journal: Bio-protocol

Article Title: In Ovo CAM-Based Xenograft Model for Investigating Tumor Developmental Biology in Breast Cancer

doi: 10.21769/BioProtoc.5600

Figure Lengend Snippet: Macroscopic images of in ovo tumors showing (A) MCF-7, characterized by a compact cellular mass with regular edges, and (B) MDA-MB-231, displaying a tumor with irregular, radially shaped edges. The vascular network of the CAM is visible beneath the tumor mass. Scale bars, 2 mm. (C–D) Panoramic views of hematoxylin-eosin-stained histological sections of the tumors developed in the CAM model: (C) MCF-7 and (D) MDA-MB-231; scale bars, 1 mm. (E–F) Higher-magnification views of the regions indicated by the red boxes in panels C and D: (E) MCF-7 and (F) MDA-MB-231. In both tumors, areas of neoplastic cells with a denser staining intensity embedded within the stroma are evident. Scale bars, 300 μm. Arrows indicate blood vessels lined by endothelial cells and containing red blood cells, present in both tumors.

Article Snippet: Cell line MCF-7 (ATCC ® HTB-22) 3.

Techniques: In Ovo, Staining