normal human lung fibroblast cells imr90 ccl 186 atcc united states of america  (ATCC)

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: A. Schema of IMR90 senescence induced by etoposide. B. SA-β-gal staining of senescent IMR90 fibroblasts at 3 weeks post-treatment with etoposide (right panel). 200x magnification, scale bar represents 100 µm. C. Cytokine profiles of analytes that decreased post- etoposide treatment in IMR90 cells. D. Cytokine profiles of analytes that increased post-etoposide treatment in IMR90 cells. Blue circles represent pre-etoposide analyte concentrations in picograms per milliliter (pg/mL), red squares represent analytes concentrations 1-week post- etoposide, and purple inverted triangles represent analyte concentrations 3-weeks post- etoposide.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques: Staining

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: Senescent fibroblasts promote tumor cell proliferation in co-culture and tumor growth in vivo .. A. Schema of co-culture system with constitutive Luciferase reporter expression in HT29 cancer cells. B. Relative bioluminescence in a two-cell co-culture system using HT29 tumor cells and IMR90 fibroblasts. Relative bioluminescence was normalized to that of HT29-luc cocultured with senescent IMR90 at each day indicated. C. Imaging of tumor cells in co-culture system at day 6. Scale bar represents 100 microns. D. Bioluminescence in co-culture system at 3 and 6 days post-ABT263 treatment. E. Bioluminescence in co-culture system at 3 and 6 days post-3TC treatment. F. Heat map representing cytokine, chemokine, and growth factor profiles. Fold changes post-treatment with ABT-263 (5µM) or 3TC (10µM) are shown in both senescent and proliferating IMR90s. G. HT29 mouse xenograft average tumor growth volumes on day 5 after HT29 cells were subcutaneously implanted with senescent or proliferating IMR90 cells. H. Tumor growth curves of HT29 CRC xenografts in mice. Tumor growth is reported according to tumor size measurements. Tumor volumes (G and H) were measured by a caliper every three days. Relative bioluminescence was normalized to control in each cohort, respectively. Data are expressed as mean ± SD. *, p <0.05.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques: Co-Culture Assay, In Vivo, Luciferase, Expressing, Imaging, Control

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: HCT116-Luc cells were cocultured with the senescent IMR90 or proliferating MR90 cells. Relative bioluminescence was normalized to the control in each cohort, respectively. Data are expressed as mean ± SD. *, p <0.05.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques: Control

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: A. Cell viability graph representing senescent and proliferating IMR90 cell viability post treatment with ABT263 at indicated concentrations. B. Cell viability graph representing senescent and proliferating IMR90 cell viability post treatment with 3TC at indicated concentrations.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques:

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: A. Schema of culture of p21-knockdown senescent IMR90 with bystander cancer cells. B. Bioluminescence of bystander cells in two-cell co-culture system on day 3. C. SA-β-gal staining of p21-Knockdown senescent IMR90 (lower panel). p21 expression in p21-knockdown or p53-knockdown senescent IMR90 cells is shown (upper panel). D. Heat map representing cytokine, chemokine, and growth factor profiles. Fold changes are shown for p21-knockdown senescent IMR90 cells. Relative cytokine levels were normalized to those of si-Ctrl in senescent IMR90 cells. Relative cytokine levels from senescent IMR90 with si-Ctrl were normalized to those from proliferating IMR90 cells. E. Bioluminescence of bystander HT29 cancer cells in two-cell co- culture system with TNFα- knockdown senescent IMR90 cells. Data are expressed as mean ± SD. #, p≤0.1. F. Western blot assay for TNFα and p21 expression in senescent IMR90 cells (E). G. Schematic representation of senescent fibroblasts promote bystander cancer cell growth via p21-driven SASP in the TME. Relative bioluminescence was normalized to control for each cohort, respectively. Data are expressed as mean ± SD. *, p <0.05.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques: Knockdown, Co-Culture Assay, Staining, Expressing, Western Blot, Control

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: A. Bioluminescence of HT29-luc cells in two-cell co-culture system treated with TRAIL and TIC10/ONC201 as indicated. Relative bioluminescence was normalized to control for each cohort, respectively. B. Cell viability of IMR90 fibroblasts treated with TRAIL and ONC201. C. HT29 CRC colony formation using co-culture system treated with 3 µM of ONC201 or 100 ng/ml of TRAIL. D. HT29 colony formation in co-culture system treated with 5-FU (1 µg/ml). E. Imaging of HT29 CRC colony formation (quantified in panels C and D). F. Heat map representing cytokine, chemokine, and growth factor profiles. Fold-changes post-treatment with ONC201 (5 µM) or ONC212 (5 µM) are shown for both senescent and proliferating IMR90s. Data are expressed as mean ± SD. *, p <0.05. P+H, HT29 cells were cocultured with the proliferating IMR90 cells; S+H, HT29 cells were cocultured with senescent IMR90 cells.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques: Co-Culture Assay, Control, Imaging

Journal: bioRxiv

Article Title: TRAIL pathway suppression of cancer cell growth and immune cell-mediated tumor cell-killing in a senescent fibroblast-constructed tumor microenvironment

doi: 10.1101/2023.11.30.569479

Figure Lengend Snippet: A. Tri-culture of HT29 CRC cells (blue), senescent IMR90 fibroblasts (green), and NK-92 cells (unlabeled). B. Tri-culture of HT29 CRC cells (blue), proliferating IMR90 fibroblasts (green), and NK-92 cells (unlabeled) at 8-hour post- treatment timepoint. C. Quantification of senescent IMR90 + HT29 CRC cells + NK-92 tri-culture (panel A). D. Quantification of proliferating IMR90 + HT29 + NK-92 tri-culture (panel B). E. Quantification of senescent IMR90 + HT29 + TALL-104 tri-culture (panel G). F. Quantification of proliferating IMR90 + HT29 + TALL-104 tri-culture (panel H). G. Tri-culture of HT29 tumor cells (blue), senescent IMR90 fibroblasts (green), and TALL-104 T cells (unlabeled). H. Tri-culture of HT29 tumor cells (blue), proliferating IMR90 fibroblasts (green), and TALL-104 T cells (unlabeled). Ethidium homodimer (EthD-1) was used to visualize dead cells, 10x magnification, scale bar indicates 100 μm. One-way Anova followed by post-hoc Tukey’s multiple comparisons test was used to determine statistical significance at p < 0.05. The following symbols * and ** represent, p < 0.05 and p < 0.01, respectively. I. TALL-104 cell Granzyme B secretion post-ABT263 or ONC201 treatment was quantified using an IsoPlexis innate-immune chip single-cell cytokine profiling assay. J. A Uniform Manifold Approximation and Projection (UMAP) plot was used to cluster the single TALL-104 cells post-treatment as indicated using IsoPlexis technology. The cells were treated with ABT263 (2 µM) and ONC201 (2 µM), or the agents alone.

Article Snippet: Cell lines used in this study include human colorectal adenocarcinoma HT-29 cells (ATCC), normal lung fibroblast IMR90 cells (ATCC), human CD8+ cytotoxic TALL-104 cells (ATCC), and human natural killer NK-92 cells (previously provided by Dr. Kerry Campbell at Fox Chase Cancer Center).

Techniques:

Journal: Molecules and Cells

Article Title: Radiation-Induced CXCL12 Upregulation via Histone Modification at the Promoter in the Tumor Microenvironment of Hepatocellular Carcinoma

doi: 10.14348/molcells.2019.2280

Figure Lengend Snippet: (A) Transcript levels of CXCL12-CXCR4/7 axis genes were elevated in the heterotopic tumor model after radiation. mRNA levels of CXCL12-CXCR4/7 genes were measured approximately 1 or 3 days after exposure to radiation. The mRNA level of the control sample without radiation was arbitrarily set to one. (B) In the human hepatoma xenograft model, radiation enhanced the CXCL12-CXCR4/7 genes. After irradiation, the mRNA levels were measured 3 to 5 days later in the xenografted Huh7 cells. The levels of the unirradiated samples were arbitrarily set to one. (C) Expression of CXCL12-CXCR4/7 pathway genes increased 3 to 5 days after radiation in human hepatoma Huh7 cells cocultured with IMR90 cells. (D) The coculture (Huh7 + IMR90) effect on the radiation-induced CXCL12 expression was analyzed compared to the single-cultured Huh7 cells. Huh7 cells were grown in single- (Huh7 only) or coculture with IMR90 cells (Huh7 + IMR90) and exposed to radiation (IR) or not. The CXCL12 mRNA levels in the unirradiated single- or cocultured Huh7 cells were set to one. (E) The radiation or coculture-induced CXCL12 proteins were evaluated using ELISA. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The human hepatocarcinoma cell line Huh7, normal lung fibroblast cell line IMR90, and WI38 cells (purchased from the American Type Culture Collection) were cultured in DMEM (Welgene, Korea) supplemented with 10% fetal bovine serum (FBS; HyClone, USA) and 1% penicillin/streptomycin (Gibco, USA).

Techniques: Control, Irradiation, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Molecules and Cells

Article Title: Radiation-Induced CXCL12 Upregulation via Histone Modification at the Promoter in the Tumor Microenvironment of Hepatocellular Carcinoma

doi: 10.14348/molcells.2019.2280

Figure Lengend Snippet: The levels of the unirradiated samples were arbitrarily set to one. (A) At the murine CXCL12 promoter, alterations in trimethylation at histone 3 lysine 4 (H3K4me3) and lysine 9 (H3K9me3) were monitored 1 to 3 days after radiation. (B) Modifications of H3K4me3, H3K9me3, and acetylation at histone 3 lysine 9 residue (H3K9ac) at the human CXCL12 promoter were evaluated 1 to 3 days after radiation in the xenografted Huh7 cells. (C and D) The experiment in (B) was repeated in single-cultured (C) and cocultured (D) Huh7 cells with or without radiation. Huh7 cells were cultured alone (C; Huh7) or together with IMR90 cells (D; Huh7 co-cultured with IMR90) and treated with or without γ-irradiation. CXCL12 mRNA levels were analyzed 3 or 5 days after irradiation. (E) Huh7 cells were treated with etoposide or phleomycin for 2 h. Three days after treatment with DNA damaging agents, CXCL12 mRNA levels were analyzed and modifications of H3K4me3 and H3K9me3 at the human CXCL12 promoter were evaluated. (F) Huh7 cells were pretreated with 10 μM ATM inhibitor (KU 55933; Sigma) for 30 min, followed by irradiation, CXCL12 mRNA levels were monitored. Modifications in H3K4me3 and H3K9me3 at the human CXCL12 promoter were measured. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The human hepatocarcinoma cell line Huh7, normal lung fibroblast cell line IMR90, and WI38 cells (purchased from the American Type Culture Collection) were cultured in DMEM (Welgene, Korea) supplemented with 10% fetal bovine serum (FBS; HyClone, USA) and 1% penicillin/streptomycin (Gibco, USA).

Techniques: Residue, Cell Culture, Irradiation

Journal: Molecules and Cells

Article Title: Radiation-Induced CXCL12 Upregulation via Histone Modification at the Promoter in the Tumor Microenvironment of Hepatocellular Carcinoma

doi: 10.14348/molcells.2019.2280

Figure Lengend Snippet: (A) Recombinant CXCL12 (rCXCL12) induces expression of CXCL12, but AMD3100 inhibits the rCXCL12-induced CXCL12 expression in the single- (Huh7) and cocultured (Huh7 + IMR90) Huh7 cells. Huh7 cells were cultured alone (Huh7) or together with IMR90 cells (Huh7 + IMR90) and treated with rCXCL12 or AMD3100 for 24 to 72 h as described. CXCL12 mRNA levels were analyzed. (B) Modifications of H3 at CXCL12 promoters by rCXCL12 or AMD3100 were analyzed in Huh7 cells for 24 to 72 h after treatment with rCXCL12 or AMD3100. ChIP assays were performed using anti-H3K4me and H3K9me antibodies. (C and D) The experiments in (A) were repeated for CXCR4 (C) and CXCR7 expression (D). * P < 0.05, ** P < 0.01, *** P < 0.001. (E) Huh7 cells transfected with an empty (mock) or CXCL12 silencing (shCXCL12) vectors one day earlier then irradiation or phleomycin treatment. Scale bar = 100 μm. Invasion was evaluated 3 days after irradiation or phleomycin treatment by incubating Huh7 cells on a Matrigel-coated chamber.

Article Snippet: The human hepatocarcinoma cell line Huh7, normal lung fibroblast cell line IMR90, and WI38 cells (purchased from the American Type Culture Collection) were cultured in DMEM (Welgene, Korea) supplemented with 10% fetal bovine serum (FBS; HyClone, USA) and 1% penicillin/streptomycin (Gibco, USA).

Techniques: Recombinant, Expressing, Cell Culture, Transfection, Irradiation

Journal: Molecules and Cells

Article Title: Radiation-Induced CXCL12 Upregulation via Histone Modification at the Promoter in the Tumor Microenvironment of Hepatocellular Carcinoma

doi: 10.14348/molcells.2019.2280

Figure Lengend Snippet: (A) Huh-7 cells were cultured with (cocultured) or without IMR90 cells and pretreated with IOX-1 inhibitor for 24 h followed by IR exposure. The CXCL12 mRNA level of the unpretreated and unirradiated sample was arbitrarily set to one and compared to treated samples. (B and C) Expression of CD133 + CSC marker was also affected by inhibitors after treatment with IR. (D–F) Huh-7 cells were pretreated with IOX-1 inhibitor for 24 h before irradiation. (D) In 3 days, modifications of H3K4me3 and H3K9me3 at the human CXCL12 promoter were evaluated. (E) MMP2 and MMP9 mRNA were measured. * P < 0.05, ** P < 0.01, *** P < 0.001. (F) Invasion was evaluated by incubating Huh7 cells in a Matrigel-coated chamber. Scale bar = 100 μm.

Article Snippet: The human hepatocarcinoma cell line Huh7, normal lung fibroblast cell line IMR90, and WI38 cells (purchased from the American Type Culture Collection) were cultured in DMEM (Welgene, Korea) supplemented with 10% fetal bovine serum (FBS; HyClone, USA) and 1% penicillin/streptomycin (Gibco, USA).

Techniques: Cell Culture, Expressing, Marker, Irradiation

Journal: PLoS ONE

Article Title: Antitumor Activity of a 5-Hydroxy-1 H -Pyrrol-2-(5 H )-One-Based Synthetic Small Molecule In Vitro and In Vivo

doi: 10.1371/journal.pone.0128928

Figure Lengend Snippet: (A) The chemical structure of 1d . (B) Cells were treated with 1d at 5 or 10 μg/ml or with DMSO (control) for 48 h and were analyzed using the MTT assay. Data are shown as the means of 2 independent experiments ± SEM. (C) The indicated cells were treated with 1d (1 μg/ml) or DMSO for the indicated days and stained with crystal violet. (D) HCT116 and IMR90 cells were treated with a serial diluted 1d for 24 h (left) and 48 h (right) and were analyzed using the MTT assay.

Article Snippet: Human colorectal carcinoma cell line HCT116, cervix adenocarcinoma cell line HeLa, osteosarcoma cell line U2OS, non-small cell lung cancer cell line H1299, liver hepatocellular carcinoma cell line HepG2 and normal human lung fibroblasts cell line IMR90 were obtained from the American Type Culture Collection (ATCC).

Techniques: Control, MTT Assay, Staining

Journal: PLoS ONE

Article Title: Antitumor Activity of a 5-Hydroxy-1 H -Pyrrol-2-(5 H )-One-Based Synthetic Small Molecule In Vitro and In Vivo

doi: 10.1371/journal.pone.0128928

Figure Lengend Snippet: (A-C) HCT116 cells treated with control (DMSO), Dox or 1d were analyzed using Western blot using the indicated antibodies (A and C) or were subjected to real-time PCR analysis (B). Data are the means of 2 independent experiments ± SEM. *, p<0.05; **, p<0.005; ***, p<0.001. (D and E) HCT116, H1299, U2OS and IMR90 cells treated with DMSO, Dox or 1d were immunostained followed by fluorescence microscopy (D) or were analyzed using Western blot using the indicated antibodies (E).

Article Snippet: Human colorectal carcinoma cell line HCT116, cervix adenocarcinoma cell line HeLa, osteosarcoma cell line U2OS, non-small cell lung cancer cell line H1299, liver hepatocellular carcinoma cell line HepG2 and normal human lung fibroblasts cell line IMR90 were obtained from the American Type Culture Collection (ATCC).

Techniques: Control, Western Blot, Real-time Polymerase Chain Reaction, Fluorescence, Microscopy