Journal: Nutrients

Article Title: Essential Oils, Pituranthos chloranthus and Teucrium ramosissimum , Chemosensitize Resistant Human Uterine Sarcoma MES-SA/Dx5 Cells to Doxorubicin by Inducing Apoptosis and Targeting P-Glycoprotein

doi: 10.3390/nu13051719

Figure Lengend Snippet: Effects of Pituranthos chloranthus (PC) and Teucrium ramosissimum Desf. (TR) extracts on normal primary human uterine fibroblast cells (HUF) and primary murine Bone Marrow-Derived Macrophages (BMDM) viability. After treatment of primary HUF and murine BMDM with increasing concentrations (0–100 µg/mL) of PC and TR for 72 h, the percentage of viable cells was assessed using the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. ( A ) Dose–response curves of PC-treated HUF (left panel) and TR-treated HUF (right panel). ( B ) Dose–response curves of PC-treated BMDM (left panel) and TR-treated BMDM (right panel). Data are expressed as a mean percentage of control growth ± Standard Deviation (SD) of two representative experiments ( n = 6 replicates per concentration).

Article Snippet: Primary human uterine fibroblast normal cells (HUF) were obtained from the ATCC.

Techniques: Derivative Assay, MTT Assay, Control, Standard Deviation, Concentration Assay

Journal: The Journal of investigative dermatology

Article Title: Wavelength-specific synergy between ultraviolet radiation and interleukin-1 alpha in the regulation of matrix-related genes: mechanistic role for tumor necrosis factor-alpha.

doi: 10.1046/j.1523-1747.1999.00681.x

Figure Lengend Snippet: Figure 1. Effects of specific wavelengths on the ability of IL-1α to stimulate matrix-related mRNA in fibroblasts. Human cultured fibroblasts were irradiated with 30 mJ of UVB, 5 J of UVA, 5 J of UVA1, or sham irradiated, and then given media without (minus symbols) or with (plus symbols) 10 ng IL-1α per ml, followed by 24 h of incubation at 37°C. Displayed are normalized levels of tropoelastin (A) and collagenase (B) mRNA.

Article Snippet: Cultured cells Normal human fibroblasts were obtained from the American Type Culture Collection (ATCC, Rockville, MD, catalog no. 1828X-CRL) and grown in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum.

Techniques: Cell Culture, Irradiation, Incubation

Journal: The Journal of investigative dermatology

Article Title: Wavelength-specific synergy between ultraviolet radiation and interleukin-1 alpha in the regulation of matrix-related genes: mechanistic role for tumor necrosis factor-alpha.

doi: 10.1046/j.1523-1747.1999.00681.x

Figure Lengend Snippet: Figure 2. Effects of specific wavelengths on cellular secretion of TNF-α. (a) Fibroblasts were irradiated with 30 mJ of UVB, 5 J of UVA, 5 J of UVA1, or sham irradiated, and then given media without (minus symbols) or with (plus symbols) 10 ng IL-1α per ml, followed by 24 h of incubation at 37°C. Displayed are TNF-α concentrations in the conditioned media (n 5 2). (b) Keratinocytes were similarly irradiated, incubated, and media assayed for TNF-α. Displayed are TNF-α concentrations in the conditioned media when no exogenous IL-1α had been added (n 5 2). Separate experiments demonstrated that exogenous IL-1α did not affect secretion of TNF-α by keratinocytes exposed to any of these wavelengths. (c) Neonatal and adult keratinocytes were irradiated with 10 or 30 mJ per cm2 of UVB or 5 or 20 J per cm2 of UVA. Displayed are TNF-α concentrations in the conditioned media from irradiated neonatal (n 5 2) and adult (n 5 3) keratinocytes, p , 0.01 for adult 30 mJ per cm2 UVB relative to sham by analysis of variance, then Dunnett q9 statistic.

Article Snippet: Cultured cells Normal human fibroblasts were obtained from the American Type Culture Collection (ATCC, Rockville, MD, catalog no. 1828X-CRL) and grown in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum.

Techniques: Irradiation, Incubation

Journal: The Journal of investigative dermatology

Article Title: Wavelength-specific synergy between ultraviolet radiation and interleukin-1 alpha in the regulation of matrix-related genes: mechanistic role for tumor necrosis factor-alpha.

doi: 10.1046/j.1523-1747.1999.00681.x

Figure Lengend Snippet: Figure 3. Effects of exogenous TNF-α on the ability of IL-1α to stimulate tropoelastin mRNA in fibroblasts. Human cultured fibroblasts were incubated with or without IL-1α and TNF-α, as indicated. Displayed are normalized tropoelastin mRNA levels after 24 h of incubation at 37°C. IL-1α produced a statistically significant increase in tropoelastin mRNA only in the cells that received no TNF-α (p , 0.01 by t test).

Article Snippet: Cultured cells Normal human fibroblasts were obtained from the American Type Culture Collection (ATCC, Rockville, MD, catalog no. 1828X-CRL) and grown in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum.

Techniques: Cell Culture, Incubation, Produced

Journal: The Journal of investigative dermatology

Article Title: Wavelength-specific synergy between ultraviolet radiation and interleukin-1 alpha in the regulation of matrix-related genes: mechanistic role for tumor necrosis factor-alpha.

doi: 10.1046/j.1523-1747.1999.00681.x

Figure Lengend Snippet: Figure 4. Effect of anti-TNF-α antibodies on the ability of UVB to block the effect of IL-1α on matrix-related messages. Human cultured fibroblasts were irradiated with 30 mJ of UVB or sham irradiation, and then given media without (minus symbols) or with (plus symbols) 10 ng IL-1α per ml, plus the indicated concentrations of anti-TNF-α inhibitory antibodies. Displayed are normalized tropoelastin (a) and collagen (b) mRNA levels after 24 h of incubation at 37°C.

Article Snippet: Cultured cells Normal human fibroblasts were obtained from the American Type Culture Collection (ATCC, Rockville, MD, catalog no. 1828X-CRL) and grown in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum.

Techniques: Blocking Assay, Cell Culture, Irradiation, Incubation

Journal: Journal of Biological Chemistry

Article Title: Evidence That Factor VIII Forms a Bivalent Complex with the Low Density Lipoprotein (LDL) Receptor-related Protein 1 (LRP1)

doi: 10.1074/jbc.m116.754622

Figure Lengend Snippet: FIGURE 7. Effect of clusters II and IV on LRP1-mediated internalization of BDD-fVIII (A) and tPAPAI-1 (B) by WI-38 fibroblasts. WI-38 fibroblasts were seeded in 12-well tissue culture dishes (1 105 cells/well) and incubated with 20 nM 125I-BDD-fVIII (A) or 10 nM 125I-tPAPAI-1 (B) for 4 h at 37 °C in the presence orabsenceofLRP1clusterIIorIV(500nM).Followingincubation,theamountofradiolabeledligandinternalizedwasdetermined,andthedatawerenormalized to the amount of fVIII internalized in the absence of competitors. The results represent the mean of three independent experiments. Error bars represent S.E. (*, p 0.002, one-way analysis of variance with Tukey’s multiple comparison post-test).

Article Snippet: WI38 cells, human lung fibroblast cells, were obtained from American Type Culture Collection (ATCC) and maintained in DMEM (Corning) supplemented with 10% FBS and penicillin/streptomycin.

Techniques: Incubation, Comparison

Journal: Translational Oncology

Article Title: Epithelial Ovarian Cancer-Induced Angiogenic Phenotype of Human Omental Microvascular Endothelial Cells May Occur Independently of VEGF Signaling 1 2

doi:

Figure Lengend Snippet: Ovarian cancer cell secretome increases HOMEC cellular responses and contains a range of potential proangiogenic proteins. (A) Proliferation of HOMECs treated with CM from SKOV3 and A2780 cells for 24, 48, or 72 hours assessed using the WST-1 assay. Controls received basal medium only. **P ≤ .01 and ***P ≤ .001 versus control levels (100%). (B) Migration of HOMECs in co-culture with ovarian cancer cells and HDFs (co-culture control) assessed by the ThinCert migration assay. After 24 hours, fluorescence of migrated cells was quantified. ***P ≤ .001 versus control (100%) and ###P ≤ .001 versus HDFs. (C) HOMEC Oris cell migration after 48 hours in the presence of VEGFA165 (20 ng/ml) ± SU5416 (10 µM). Controls received medium alone. ***P ≤ .001 versus control (100%) and ###P ≤ .001 versus VEGFA165. (D) Migration of HOMECs in co-culture with ovarian cancer cells ± anti-VEGFA antibody (500 ng/ml) or SU5416 (10 µM) assessed by the ThinCert migration assay. ***P ≤ .001 versus control (100%), **P ≤ .01 versus control (100%), and NS versus A2780 or SKOV3. (E) VEGF- and EOC cell-induced tube-like structure formation of HOMECs and the effects of the SU5416 inhibitor. HOMECs were plated onto fibrin matrices and exposed to either VEGFA165 or EOC cells in co-culture ± SU5416 (10 µM). Negative controls received medium alone. Tube-like structure formation was quantified. *P ≤ .05, **P ≤ .01, and ***P ≤ .001 versus control (100%), ###P ≤ .001 versus VEGF, and NS versus SKOV3/A2780. For A to E, data are presented as means ± SD. P values were calculated by one-way analysis of variance (ANOVA), followed by Tukey's post hoc test. (F) Shotgun proteomics of SKOV3 CM by the 4800 MALDI TOF/TOF analyzer. Significant results were determined by selecting proteins that matched with two or more peptides with total ion CIs of >95%, followed by a literature search to determine proteins with a possible role in angiogenesis. (G) Analysis of CL, CD, IGFBP-7, and VEGF concentration in CM from SKOV3 and A2870. Commercially available ELISAs were used for determination of VEGF, CD, and CL concentrations, whereas the IGFBP-7 ELISA protocol was developed in-house. All experiments were carried out in duplicate on three separate samples. B/D, below detection limit.

Article Snippet: Ovarian cancer cell lines, SKOV3 and A2780, and human dermal fibroblasts (HDFs) were purchased from the European Collection of Cell Cultures (ECACC; Salisbury, United Kingdom) and PromoCell (Heidelberg, Germany), respectively.

Techniques: WST-1 Assay, Control, Migration, Co-Culture Assay, Fluorescence, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Oncology

Article Title: Silencing SAPCD2 Represses Proliferation and Lung Metastasis of Fibrosarcoma by Activating Hippo Signaling Pathway

doi: 10.3389/fonc.2020.574383

Figure Lengend Snippet: SAPCD2 is upregulated in fibrosarcoma tissues and cells. (A) Real-time PCR and Western blotting analysis of SAPCD2 expression in 4 normal fibroblast cell lines, including human dermal fibroblast primary cell (hDFPC), human lung fibroblasts primary cell (hLFPC), human mammary fibroblasts primary cell (hMFPC), Human embryonic lung fibroblast (HFL1), and 2 fibrosarcoma cell lines HT-1080 and SW684. GAPDH was used as endogenous controls in RT-PCR and α-Tubulin was detected as a loading control in the Western blot. Each bar represents the mean values ± SD of three independent experiments. *P < 0.05. (B) Representative images of SAPCD2 expression in benign fibroma, dermatofibrosarcoma protuberans (DFSP), and fibrosarcoma tissues. (C) The number of fibroma, DFSP, and fibrosarcoma tissues stratified by staining index of IHC. (D) Immunohistochemical staining index of SAPCD2 in fibroma, DFSP, and fibrosarcoma tissues. n.s. indicates no significance.

Article Snippet: The normal fibroblast cells human dermal fibroblast primary cell (hDFPC), human lung fibroblasts primary cell (hLFPC), human mammary fibroblasts primary cell (hMFPC), human embryonic lung fibroblast (HFL1), and one fibrosarcoma cell HT-1080 were obtained from Procell (Wuhan, China).

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Staining, Immunohistochemical staining

Journal: Frontiers in Oncology

Article Title: Silencing SAPCD2 Represses Proliferation and Lung Metastasis of Fibrosarcoma by Activating Hippo Signaling Pathway

doi: 10.3389/fonc.2020.574383

Figure Lengend Snippet: SAPCD2 inactivates Hippo signaling pathway. (A) GSEA analysis showed that SAPCD2 expression level was associated with the Hippo signaling. (B) TEAD transcriptional activity was assessed by HOP-Flash luciferase reporter in the indicated cells. Error bars represent the mean ± S.D. of three independent experiments. * P < 0.05 . (C) Western blot analysis of total p-MST1/2, MST1, p-LATS1, LATS1, p-YAP1, YAP1 expression and nuclear YAP, TAZ expression in the indicated cells. α-Tubulin was detected as a loading control and nuclear protein p84 was used as the nuclear protein marker. (D) Real-time PCR analysis of CTGF, CYR61, SOX9, HOXA1, RPL13A, and PPIA in the indicated cells. Error bars represent the mean ± S.D. of three independent experiments. * P < 0.05 . (E) Western blotting analysis of SAPCD2 expression in 4 normal fibroblast cell lines, including human dermal fibroblast primary cell (hDFPC), human lung fibroblasts primary cell (hLFPC), human mammary fibroblasts primary cell (hMFPC), Human embryonic lung fibroblast (HFL1), and 2 fibrosarcoma cell lines HT-1080 and SW684. α-Tubulin was detected as a loading control.

Article Snippet: The normal fibroblast cells human dermal fibroblast primary cell (hDFPC), human lung fibroblasts primary cell (hLFPC), human mammary fibroblasts primary cell (hMFPC), human embryonic lung fibroblast (HFL1), and one fibrosarcoma cell HT-1080 were obtained from Procell (Wuhan, China).

Techniques: Expressing, Activity Assay, Luciferase, Western Blot, Control, Marker, Real-time Polymerase Chain Reaction