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: Oncology Reports

Article Title: Filamin B extensively regulates transcription and alternative splicing, and is associated with apoptosis in HeLa cells

doi: 10.3892/or.2020.7532

Figure Lengend Snippet: Knockdown of FLNB promotes proliferation and inhibits apoptosis of HeLa cells. (A) Relative mRNA expression of FLNB in HeLa cells after it was knocked down using FLNB -specific shRNA was determined by reverse transcription-qPCR. shFLNB_1 and shFLNB_2 indicate two biological repeats of HeLa cells transfected with FLNB-specific shRNA. Ctrl_1 and Ctrl_2 indicate two biological repeats of HeLa cells transfected with empty vector as controls. (B) Cell proliferation of shFLNB was measured by an MTT assay in HeLa cells. Cell apoptosis of cells transfected with shFLNB was measured by (C) flow cytometry and (D) subsequent analysis by a 7-ADD and Annexin V assay. *P<0.05 and **P<0.01 vs. respective Ctrl. FLNB , filamin B; sh, short hairpin; qPCR, quantitative PCR; Ctrl, control; 7-AAD, 7-amino actinomycin D; OD, optical density; PE, phycoerythrin.

Article Snippet: Human HeLa cells (CCTCC@GDC0009) were obtained from The China Center for Type Culture Collection.

Techniques: Knockdown, Expressing, shRNA, Reverse Transcription, Transfection, Plasmid Preparation, MTT Assay, Flow Cytometry, Annexin V Assay, Real-time Polymerase Chain Reaction, Control

Journal: Oncology Reports

Article Title: Filamin B extensively regulates transcription and alternative splicing, and is associated with apoptosis in HeLa cells

doi: 10.3892/or.2020.7532

Figure Lengend Snippet: GO and KEGG analysis of differentially expressed genes between short hairpin FLNB-transfected and control HeLa cells. Top 10 most enriched GO terms (biological process) of (A) upregulated and (B) downregulated genes upon FLNB knockdown. Rectangles around GO terms indicate notable cancer-related and cartilage development terms. Top 10 most enriched KEGG pathways of (C) upregulated and (D) downregulated genes upon FLNB knockdown. GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; FLNB , filamin B.

Article Snippet: Human HeLa cells (CCTCC@GDC0009) were obtained from The China Center for Type Culture Collection.

Techniques: Transfection, Control, Knockdown

Journal: Oncology Reports

Article Title: Filamin B extensively regulates transcription and alternative splicing, and is associated with apoptosis in HeLa cells

doi: 10.3892/or.2020.7532

Figure Lengend Snippet: Validation of FLNB -regulated genes (DEGs). (A) Relative expression level (FPKM, up) and RT-qPCR measurement (down) of cartilage development-related DEGs. (B) Related expression level (FPKM, up) and RT-qPCR measurement (down) of apoptotic-related DEGs. (C) Western blot analysis of two apoptotic-related proteins in shFLNB and Ctrl HeLa cells. FLNB , filamin B; DEGs, differentially expressed genes; FPKM, fragments per kilobase of transcript per million fragments mapped; RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin; Ctrl, control; ATP7A, ATPase copper transporting α; BMP7, bone morphogenetic protein 7; COL2A1, collagen type II α 1 chain; MMP13, matrix metallopeptidase 13; IL23A, interleukin 23 subunit α; MALAT1, metastasis associated lung adenocarcinoma transcript 1; NAIP, NLR family apoptosis inhibitory protein; SLC25A36, solute carrier family 25 member 36; MAP2K7, mitogen-activated protein kinase kinase 7.

Article Snippet: Human HeLa cells (CCTCC@GDC0009) were obtained from The China Center for Type Culture Collection.

Techniques: Biomarker Discovery, Expressing, Quantitative RT-PCR, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Control

Journal: Oncology Reports

Article Title: Filamin B extensively regulates transcription and alternative splicing, and is associated with apoptosis in HeLa cells

doi: 10.3892/or.2020.7532

Figure Lengend Snippet: Validation of FLNB -affected ASEs. Genome visualization (left panel) shows FLNB -regulated ASEs in shFLNB and control. (A) Validation of an ASE of VDR in HeLa cells. (B) Validation of an ASE of PACS2 in HeLa cells. (C) Validation of an ASE of MAP2K7 in HeLa cells. (D) Validation of an ASE of MALAT1 in HeLa cells. The number of junction reads were marked on the line representing splice junction composing ASE. The structures of ASEs are depicted in the top-right panel. The altered ratio of ASEs in RNA-sequencing and in reverse transcription-quantitative PCR were calculated and plotted (right panel, bottom). FLNB , filamin B; ASEs, alternative splicing events; sh/SH, short hairpin; NC, negative control; VDR, vitamin D receptor; PACS2, phosphofurin acidic cluster sorting protein 2; MAP2K7, mitogen-activated protein kinase kinase 7; MALAT1, metastasis associated lung adenocarcinoma transcript 1. *P<0.05 and ***P<0.001 vs. respectively NC.

Article Snippet: Human HeLa cells (CCTCC@GDC0009) were obtained from The China Center for Type Culture Collection.

Techniques: Biomarker Discovery, Control, RNA Sequencing, Reverse Transcription, Real-time Polymerase Chain Reaction, Alternative Splicing, Negative Control

Journal: International Journal of Biological Sciences

Article Title: E2F8 regulates the proliferation and invasion through epithelial-mesenchymal transition in cervical cancer

doi: 10.7150/ijbs.37686

Figure Lengend Snippet: E2F8 expression in cervical cancer cell lines and correlation with suppressed cell proliferation. (A, B) The expression levels of E2F8 RNA and protein were significantly higher in some cervical cancer cell lines (CaSki, HeLa, and ME180) compared to that in normal control cells. (C) Using siE2F8, E2F8 knockdown was performed in E2F8-elevated HeLa cell lines. (D) The proliferation of HeLa cells transfected with siE2F8 and negative control siNC was determined using the CCK-8 assay. (E) Using shE2F8, E2F8 knockdown was performed in E2F8-elevated ME180 cell lines. (F) The proliferation of ME180 cells transfected with shE2F8 and negative scrambled control was determined using the CCK-8 assay. Each assay was performed in triplicate. Data are means ± standard deviation. ** P <0.01, ***P<0.001 vs siNC and scrambled control.

Article Snippet: Human epithelioid cervical adenocarcinoma (HeLa) cells were obtained from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Expressing, Control, Knockdown, Transfection, Negative Control, CCK-8 Assay, Standard Deviation

Journal: International Journal of Biological Sciences

Article Title: E2F8 regulates the proliferation and invasion through epithelial-mesenchymal transition in cervical cancer

doi: 10.7150/ijbs.37686

Figure Lengend Snippet: E2F8 promotes cell migration and invasion. (A, C) Wound healing assay observed under the optical microscope was used to determine cell migration using si and shE2F8. E2F8 knockdown was performed in E2F8-high HeLa and ME180 cell lines. (B, D) Cell invasion was observed under the optical microscope. Matrigel invasion assays were used to determine invasion after 48 h in HeLa and ME180 cells. Each assay was performed in triplicate. Data represent means ± standard deviation. ** P <0.01, *** P <0.001 vs. siNC and scrambled control.

Article Snippet: Human epithelioid cervical adenocarcinoma (HeLa) cells were obtained from the Korean Cell Line Bank (Seoul, Korea).

Techniques: Migration, Wound Healing Assay, Microscopy, Knockdown, Standard Deviation, Control