Journal: Frontiers in cell and developmental biology

Article Title: Fra-1 Inhibits Cell Growth and the Warburg Effect in Cervical Cancer Cells via STAT1 Regulation of the p53 Signaling Pathway.

doi: 10.3389/fcell.2020.579629

Figure Lengend Snippet: FIGURE 1 | Fra-1 inhibited the proliferation and promoted apoptosis of cervical cancer cells. (A) Cell viability of HeLa cells transfected with control vector (HeLa/vector) and HeLa cells transfected with Fra-1 overexpression vector (HeLa/Fra-1) cells determined by CCK8 assay. (B) Colony forming ability of HeLa/vector and HeLa/Fra-1 cells. (C) Colony numbers formed by HeLa/vector and HeLa/Fra-1 cells. (D) Cell cycle distribution among HeLa/vector and HeLa/Fra-1 cells as determined by flow cytometry. (E) Flow cytometric analysis of apoptosis among HeLa/vector and HeLa/Fra-1 cells. Data means ± standard deviation (SD). Each representative experiment was repeated three times with similar results. ∗∗P < 0.01; ∗∗∗P < 0.001, n = 3.

Article Snippet: HeLa cells, a human cervical cancer cell line, were purchased from the ATCC (Manassas, VA, United States) and maintained by our laboratory.

Techniques: Transfection, Control, Plasmid Preparation, Over Expression, CCK-8 Assay, Cytometry, Standard Deviation

Journal: Frontiers in cell and developmental biology

Article Title: Fra-1 Inhibits Cell Growth and the Warburg Effect in Cervical Cancer Cells via STAT1 Regulation of the p53 Signaling Pathway.

doi: 10.3389/fcell.2020.579629

Figure Lengend Snippet: FIGURE 2 | Fra-1 regulated p53 signaling pathway activity in cervical cancer cells. (A) mRNA chip analysis in HeLa/vector and HeLa/Fra-1 cells revealed differences in mRNA expression between the two groups. Differentially active signaling pathways were identified by gene ontology analysis. (B) Western blotting analysis of signal transducer and activator of transcription 1 (STAT1), p53, Bcl-2, p38, p21, MDM2, CDK4, and cyclinD1 protein levels in HeLa/vector and HeLa/Fra-1 cells. (C) Real-time polymerase chain reaction (PCR) analysis the mRNA expression levels of STAT1, p53, Bcl-2, p38, p21, MDM2, CDK4, and cyclinD1 in HeLa/vector and HeLa/Fra-1 cells. *P < 0.05; **P < 0.01; ***P < 0.001, n = 3.

Article Snippet: HeLa cells, a human cervical cancer cell line, were purchased from the ATCC (Manassas, VA, United States) and maintained by our laboratory.

Techniques: Activity Assay, Plasmid Preparation, Expressing, Protein-Protein interactions, Western Blot, Real-time Polymerase Chain Reaction

Journal: Frontiers in cell and developmental biology

Article Title: Fra-1 Inhibits Cell Growth and the Warburg Effect in Cervical Cancer Cells via STAT1 Regulation of the p53 Signaling Pathway.

doi: 10.3389/fcell.2020.579629

Figure Lengend Snippet: FIGURE 3 | Fra-1 promoted cell senescence and restored mitochondrial disorder in cervical cancer cells. (A) β-Galactosidase assay for cell senescence in HeLa/vector and HeLa/Fra-1 cells. (B) Western blot analysis of Fra-1, SIRT1, NF-κB, p16, and c-Myc protein expression in HeLa/vector and HeLa/Fra-1 cells. (C) Flow cytometric analysis of intracellular Ca2+ concentration. Red color represents HeLa/vector group and green color represents HeLa/Fra-1 group. (D) Flow cytometric analysis of intracellular reactive oxygen species (ROS) concentration. Red color represents HeLa/vector group and green color represents HeLa/Fra-1 group. (E) Flow cytometric analysis of mitochondrial membrane potential (1ψm) in HeLa/vector and HeLa/Fra-1 cells. (F) NAD+/NADH ratio in HeLa/vector and HeLa/Fra-1 cells. (G) Western blot analysis of sirtuin 3 (SIRT3), SOD2, IDH2, LKB1, and p-AMPK expression in HeLa/vector and HeLa/Fra-1 cells. **P < 0.01; ***P < 0.001, n = 3.

Article Snippet: HeLa cells, a human cervical cancer cell line, were purchased from the ATCC (Manassas, VA, United States) and maintained by our laboratory.

Techniques: Plasmid Preparation, Western Blot, Expressing, Concentration Assay, Membrane

Journal: Frontiers in cell and developmental biology

Article Title: Fra-1 Inhibits Cell Growth and the Warburg Effect in Cervical Cancer Cells via STAT1 Regulation of the p53 Signaling Pathway.

doi: 10.3389/fcell.2020.579629

Figure Lengend Snippet: FIGURE 5 | Silencing of signal transducer and activator of transcription 1(STAT1) by small interfering RNA promoted proliferation of cervical cancer cells via activation of the p53 signal pathway. (A) Real-time polymerase chain reaction (PCR) analysis of the silencing effects of three siRNAs for STAT1 in HeLa/Fra-1 cells. (B) Cell viability of HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells according to CCK8 assay. (C) Colony forming ability of HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (D) In vivo analysis of subcutaneous implanted tumor growth to compare the tumorigenic abilities of HeLa/vector, HeLa/Fra-1, and HeLa/Fra-1/siSTAT1 cells in mice. (E) Western blot analysis of protein expression of STAT1, CDK4, cyclin A, cyclin D1, p53, Bcl-2, c-REL, p38, p21, and MDM2 in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (F) Real-time PCR analysis of mRNA expression of STAT1, CDK4, cyclin D1, p53, Bcl-2, p38, p21, and MDM2 in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. *P < 0.05; **P < 0.01; ***P < 0.001, n = 3.

Article Snippet: HeLa cells, a human cervical cancer cell line, were purchased from the ATCC (Manassas, VA, United States) and maintained by our laboratory.

Techniques: Small Interfering RNA, Activation Assay, Real-time Polymerase Chain Reaction, CCK-8 Assay, In Vivo, Plasmid Preparation, Western Blot, Expressing

Journal: Frontiers in cell and developmental biology

Article Title: Fra-1 Inhibits Cell Growth and the Warburg Effect in Cervical Cancer Cells via STAT1 Regulation of the p53 Signaling Pathway.

doi: 10.3389/fcell.2020.579629

Figure Lengend Snippet: FIGURE 6 | Silencing of signal transducer and activator of transcription 1 (STAT1) inhibited senescence and promoted mitochondrial dysfunction in cervical cancer cells. (A) β-Galactosidase assay for cell senescence in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (B) Western blot analysis of protein expression of c-Myc, SIRT1, NF-κB, and p16 in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (C) Flow cytometric analysis of intracellular Ca2+ concentration. Red color represents HeLa/Fra-1 group and green color represents HeLa/Fra-1/siSTAT1 group. (D) Flow cytometric analysis of intracellular reactive oxygen species (ROS) levels. Red color represents HeLa/Fra-1 group and green color represents HeLa/Fra-1/siSTAT1 group. (E) MFI data reflecting changes in the intracellular ROS concentration in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (F) Flow cytometric analysis of mitochondrial membrane potential (1ψm) in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. *P < 0.05; **P < 0.01; ***P < 0.001, n = 3.

Article Snippet: HeLa cells, a human cervical cancer cell line, were purchased from the ATCC (Manassas, VA, United States) and maintained by our laboratory.

Techniques: Western Blot, Expressing, Concentration Assay, Membrane

Journal: Frontiers in cell and developmental biology

Article Title: Fra-1 Inhibits Cell Growth and the Warburg Effect in Cervical Cancer Cells via STAT1 Regulation of the p53 Signaling Pathway.

doi: 10.3389/fcell.2020.579629

Figure Lengend Snippet: FIGURE 7 | Silencing of signal transducer and activator of transcription 1(STAT1) recovered metabolic reprogramming in cervical cancer cells overexpressing Fra-1. (A) NAD + /NADH ratio in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (B) Western blot analysis of STAT1, sirtuin 3 (SIRT3), LKB1, and p-AMPK expression in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (C) Western blot analysis of the expression of key glycolysis enzymes PFK1, PKM2, and PDH in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (D) Real-time polymerase chain reaction (PCR) analysis of mRNA expression of glycolysis enzymes Glut1, HK II, and LDHA in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (E) Glucose concentration in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (F) Lactate concentration in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. (G) Protein expression of glutamine dehydrogenase (GDH) and glutamine concentration in HeLa/Fra-1 and HeLa/Fra-1/siSTAT1 cells. NS: No statistical difference; P < 0.05; **P < 0.01; ***P < 0.001, n = 3.

Article Snippet: HeLa cells, a human cervical cancer cell line, were purchased from the ATCC (Manassas, VA, United States) and maintained by our laboratory.

Techniques: Western Blot, Expressing, Real-time Polymerase Chain Reaction, Concentration Assay

Journal: bioRxiv

Article Title: Unleashing a Novel Function of Endonuclease G in Mitochondrial Genome Instability

doi: 10.1101/2021.05.27.445952

Figure Lengend Snippet: A. Representative images of HeLa and HEK293T cells showing localization of BG4, the G4 binding antibody to mitochondria following immunofluorescence study. Nucleus is stained with DAPI (blue color), mitochondria with MitoTracker DR (red) and BG4 with Alexa-Fluor 488 (green). A merged image is shown with merge of red and green as depicted by Coste’s mask (colocalization is represented as a white dot). B, C. Quantitation showing colocalization of BG4 with MitoTracker indicated as dot plots. The colocalization was quantified using Mander’s colocalization coefficient (ImageJ software) analyzing a minimum of 100 cells as red over green (B) and green over red (C). D. Representative image of rho(0) cell showing localization of BG4 to mitochondria following immunofluorescence as investigated in panel A. E. Quantitation showing comparison of colocalization of BG4 between HeLa cells and rho(0) cells is shown as dot plots. The colocalization was quantified using Mander’s colocalization coefficient analyzing a minimum of 50 cells as red over green. F. BG4 bound mtDNA was purified after reverse crosslinking and used for real-time PCR using primers derived from different regions of the mitochondrial genome, which include 5 G-quadruplex forming regions and 10 random regions. Input DNA served as template control. No antibody control was also used. Bars in blue (first 5) are for G-quadruplex forming regions, while in green (last 10) are for random regions. Y-axis depicts threshold Ct value obtained following real time PCR for each primer. Error bar represents mean ± SEM. G. Agarose gel profile showing the amplification of Input DNA (left panel) and BG4 pull down DNA (right panel). “M” denotes 100 bp ladder.

Article Snippet: HeLa (human cervical cancer), MEF (mouse embryonic fibroblast) and HEK293T (human embryonic kidney epithelial cell line) were purchased from National Centre for Cell Science, Pune, India.

Techniques: Binding Assay, Immunofluorescence, Staining, Quantitation Assay, Software, Comparison, Purification, Real-time Polymerase Chain Reaction, Derivative Assay, Control, Agarose Gel Electrophoresis, Amplification

Journal: Oncology Letters

Article Title: Identification of a peptide specifically targeting ovarian cancer by the screening of a phage display peptide library

doi: 10.3892/ol.2016.4549

Figure Lengend Snippet: Immunofluorescence assay of phage clone P2 binding HO-8910, HeLa and CHO cells. The selected phage clone P2 was added individually to the cells and then incubated with anti-M13 antibody, FITC-labeled antibody and PI. The results were visualized using a fluorescence microscope. The results show that P2 could bind to HO-8910 cells but not to HeLa and CHO cells (control cells). Magnification, ×200. PI, propidium iodide; FITC, fluorescein isothiocyanate.

Article Snippet: The HO-8910 human ovarian cancer cell line, and HeLa cervical cancer cell line, were purchased from Jilin Baili Biotechnology Co., Ltd. (Changchun, China).

Techniques: Immunofluorescence, Binding Assay, Incubation, Labeling, Fluorescence, Microscopy, Control

Journal: Cell Death & Disease

Article Title: Novel prosurvival function of Yip1A in human cervical cancer cells: constitutive activation of the IRE1 and PERK pathways of the unfolded protein response

doi: 10.1038/cddis.2017.147

Figure Lengend Snippet: Depletion of Yip1A induces apoptotic cell death in HeLa and CaSki cervical cancer cells. ( a ) HeLa and CaSki cells were transfected with control scramble siRNA (left panel) or Yip1A siRNA (right panel). Representative confocal micrographs show morphological differences between control and Yip1A-knockdown cells at the indicated time points. Scale bars are 50 μ m. ( b ) The viability of transfected cells was evaluated at the indicated time points using the MTT Cell Proliferation Assay. Data are means±S.D. from three independent experiments; * P <0.05, ** P <0.01. ( c ) Representative flow cytometric data for control (left panels) and Yip1A-knockdown (right panels) cells at the indicated time points after siRNA transfection. The percentages of apoptotic cells (Annexin V + /PI − + Annexin V + /PI + ) are shown in the bar graphs. Data are means±S.D. from three independent experiments; * P <0.05. ( d ) Representative confocal micrographs of control (upper panels) and Yip1A-knockdown (lower panels) cells at the indicated time points after siRNA transfection. Scale bars are 10 μ m. The percentages of TUNEL-positive cells are shown in the bar graphs. Data are means±S.D. from three independent experiments; * P <0.05, ** P <0.01. ( e ) Western blotting shows relative levels of cleaved caspase 3 and cleaved PARP protein in control and Yip1A-knockdown cells at the indicated time points after siRNA transfection. GAPDH was used for normalization. Data are means±S.D. from three independent experiments; * P <0.05, ** P <0.01

Article Snippet: A human cervical cancer cell line CaSki (IFO50007) was obtained from the Japanese Collection of Research Bioresources (JCRB, Osaka, Japan), and grown in RPMI 1640 Medium (Nissui) supplemented with 10% FBS (Sigma-Aldrich) and 2 mM l -glutamine (Gibco) under conditions of 5% CO 2 atmosphere at 37 °C.

Techniques: Transfection, Control, Knockdown, MTT Cell Proliferation, TUNEL Assay, Western Blot

Journal: Cell Death & Disease

Article Title: Novel prosurvival function of Yip1A in human cervical cancer cells: constitutive activation of the IRE1 and PERK pathways of the unfolded protein response

doi: 10.1038/cddis.2017.147

Figure Lengend Snippet: Schematic representation of how Yip1A operates as a prosurvival modulator that coordinately activates the IRE1 and PERK pathways of the UPR to support the survival of HeLa and CaSki cervical cancer cells. See text for details. Ub, ubiquitination

Article Snippet: A human cervical cancer cell line CaSki (IFO50007) was obtained from the Japanese Collection of Research Bioresources (JCRB, Osaka, Japan), and grown in RPMI 1640 Medium (Nissui) supplemented with 10% FBS (Sigma-Aldrich) and 2 mM l -glutamine (Gibco) under conditions of 5% CO 2 atmosphere at 37 °C.

Techniques: Ubiquitin Proteomics