Journal: PloS one

Article Title: pp32 (ANP32A) expression inhibits pancreatic cancer cell growth and induces gemcitabine resistance by disrupting HuR binding to mRNAs.

doi: 10.1371/journal.pone.0015455

Figure Lengend Snippet: Figure 6. pp32 and HuR expression in clinical samples and patient outcomes. (A) The abundance and subcellular localization of HuR (left) and low to absent nuclear pp32 expression (right) in samples from pancreatic cancer patients were assessed by immunohistochemistry; magnification, 200x. Samples are representative of the cohort analyzed in B–D. (B) Correlation between pp32 nuclear expression and response to GEM treatment (p = 0.3, log rank test). (C) Correlation between high nuclear pp32-expressing tumor samples stratified into high or low HuR status in regards to GEM response (p = 0.88, log rank test). (D) Correlation between high cytoplasmic HuR-expressing tumor samples stratified into high and low pp32 nuclear expression correlated with GEM response (p = 0.25, log rank test). doi:10.1371/journal.pone.0015455.g006

Article Snippet: MiaPaCa2 pp32-transfected (Mia.pp32) and empty vector (Mia.EV) cells were trypsinized and collected as previously described [25] and our generated do novo using the previously generated and purchased parental pancreatic cancer cell line (ATCC, Manassas, VA).

Techniques: Expressing, Immunohistochemistry

Journal: Cancer Science

Article Title: Cancer‐testis antigen BORIS is a novel prognostic marker for patients with esophageal cancer

doi: 10.1111/j.1349-7006.2012.02355.x

Figure Lengend Snippet: Expression of BORIS m RNA in various cancer cell lines and cancer tissues and presence of BORIS ‐specific IgG in sera from patients with various cancers

Article Snippet: The cell lines used in the study were esophageal squamous cell carcinoma cell lines, TE2, TE3, TE4, TE5, TE6, TE7, TE8, TE9, TE10, TE11, TE12, TE13, TE14, and TE15 (Tohoku University, Sendai, Japan); melanoma cell lines, SKmel23, SKmel28, 888mel, A375mel, 1363mel, 928mel, 624mel, 501Amel, 586mel, 526mel, 501mel, 397mel, and 1362mel (Surgery Branch, NCI, NIH, Bethesda, MD, USA); colon cancer cell line, COLO205 (JCRB, Osaka, Japan); breast cancer cell line HS578 (American Type Culture Collection (ATCC), Manassas, VA, USA); stomach cancer cell lines, MKN1, MKN7, MKN28, MKN46, and MKN74 (Yamagata University, Yamagata, Japan); endometrial cancer cell line SNGII (Keio University, Tokyo, Japan); prostate cancer cell line LNCaP (ATCC); bladder cancer cell line, KU7 (Keio University); and brain tumor cell line U87MG (ATCC).

Techniques: 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 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: Cancer research

Article Title: Inhibition of BCL2 family members increases the efficacy of copper chelation in BRAF V600E -driven melanoma

doi: 10.1158/0008-5472.CAN-19-1784

Figure Lengend Snippet: A, Schematic diagram of high throughput screen of 2123 Selleckchem bioactive compound library at 100nM alone or in combination with the IC20 of TTM (dashed red line) in A375 or WM88. B,C, Normalized Percent Inhibition (NPI) of TTM + Compound versus NPI Compound graph for indicated cells treated with Selleckchem bioactive compound library alone or in combination with IC20 of TTM. Hits (blue circles) are defined as NPI TTM + Compound ≥ 20 (dashed red line). D,E, Observed effect versus expected effect graph for indicated cells of Hits from B and C. Hits (blue circles) are defined as Bliss Index ≥ 1.5 (dashed red line). F,G, Graphical representation of Hits defined as NPI TTM + Compound ≥ 20 and Bliss Index ≥ 1.5 in indicated cells. H, Venn diagram relationship between Compound Hits. I, NPI of 5 overlapping Hit Compounds from H alone or in combination with IC20 of TTM (dashed red line). J,K, Scatter dot plot of %ATP normalized to VEH ± s.e.m. of indicated cells treated with vehicle or indicated concentrations of drugs. L,M,N,O, Scatter dot plot of %ATP normalized to VEH ± s.e.m. of indicated cells treated with VEH or indicated concentrations of drugs. Results were compared using a one-way ANOVA followed by a Tukey’s multi-comparisons test. One asterisk, P<0.05, Two asterisks, P<0.01, Three asterisks, P<0.001, Four asterisks, P<0.0001. n=3.

Article Snippet: 293T/17 (ATCC, catalog #CRL-11268), A375 (ATCC, catalog #CRL-1619), WM88 (Rockland, catalog #WM88-01-0001), WM3311 (Rockland, catalog #WM3311-01-0001), WM3743 (Rockland, catalog #WM3743-01-0001) cells were purchased from the indicated companies and maintained in Dulbecco’s Modified Eagle Media (DMEM, Gibco) supplemented with 10% v/v fetal bovine serum (FBS, GE Lifesciences) and 1% penicillin-streptomycin (P/S, Gibco).

Techniques: High Throughput Screening Assay, Drug discovery, Inhibition

Journal: Cancer research

Article Title: Inhibition of BCL2 family members increases the efficacy of copper chelation in BRAF V600E -driven melanoma

doi: 10.1158/0008-5472.CAN-19-1784

Figure Lengend Snippet: A,C, Relative CellTiter-Glo® cell viability ± s.e.m. of indicated cells stably expressing two independent Dox-inducible shRNAs (#1 and #2) against indicated genes treated with indicated concentrations of TTM without or with Dox n=3. B,D, Scatter dot plot of TTM IC50 ± s.e.m in indicated cells cells stably expressing two independent Dox-inducible shRNAs against indicated genes treated without or with Dox. Results were compared using a two-way ANOVA followed by a Sidak’s multi-comparisons test. Three asterisks, P<0.001, Four asterisks, P<0.0001. n=3. E,G, Relative CellTiter-Glo® cell viability ± s.e.m. of indicated cells treated without or with indicated concentrations of TTM and increasing concentrations of indicated BH3 mimetics. n=3. F,H, Scatter dot plot of BH3 mimetic IC50 ± s.e.m in indicated cells treated without or with indicated concentrations of TTM and increasing concentrations of indicated BH3 mimetics. n=3. Results were compared using a two-way ANOVA followed by a Tukey’s multi-comparisons test. n=3. I,J, Graphical representation of Bliss Index (observed effect versus expected effect) from A375 (E) and WM88 (G) at the indicated drug combinations. Synergistic combinations are indicated by Bliss Index values >1. Two asterisks, P<0.01, Three asterisks, P<0.001, Four asterisks, P<0.0001.

Article Snippet: 293T/17 (ATCC, catalog #CRL-11268), A375 (ATCC, catalog #CRL-1619), WM88 (Rockland, catalog #WM88-01-0001), WM3311 (Rockland, catalog #WM3311-01-0001), WM3743 (Rockland, catalog #WM3743-01-0001) cells were purchased from the indicated companies and maintained in Dulbecco’s Modified Eagle Media (DMEM, Gibco) supplemented with 10% v/v fetal bovine serum (FBS, GE Lifesciences) and 1% penicillin-streptomycin (P/S, Gibco).

Techniques: Stable Transfection, Expressing