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

Journal: Cancer Cell International

Article Title: Overexpression of trefoil factor 3 (TFF3) contributes to the malignant progression in cervical cancer cells

doi: 10.1186/s12935-016-0379-1

Figure Lengend Snippet: Forced expression of TFF3 in cervical cancer cells modulates the expression of malignant progression-related gene markers. a Determination of endogenous expression of TFF3 protein by Western blot in the cervical cancer cells lines CaSki, SiHa, Me180 and Hela and human non-tumor keratinocyte line HaCaT. b Semi-quantitative RT-PCR analysis was used to assess the mRNA levels of TFF3 in SiHa and Hela cells with either forced or depleted expression of TFF3 as described in “ ” section. c Western blot analysis was used to assess the protein levels of TFF3 in SiHa and Hela cells with either forced or depleted expression of TFF3. d , e Quantitative PCR analysis quantifying the change in expression of various genes associated with malignant progression in SiHa-TFF3/Hela cells. Change in gene expression is expressed as fold difference, respectively. Fold change values are representative of three independent experiments

Article Snippet: Human cervical cancer cell lines SiHa, CaSki, Hela, Me180 and human non-tumor keratinocyte line HaCaT were obtained from Nanjing KeyGen Biotech Co, Ltd (Nanjing,China).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Gene Expression