normal human lung fibroblast cell line imr90  (ATCC)

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

Journal: The Journal of Cell Biology

Article Title: A primate virus generates transformed human cells by fusion

doi: 10.1083/jcb.200507069

Figure Lengend Snippet: Spontaneous fusion produces proliferating hybrids. (A) IMR90 cells (I0) resistant to puromycin (I0 P ) or I0 cells transformed with E1A and resistant to hygromycin (IE H ) were cultured as indicated and stained with crystal violet to visualize the cells. (B) Co-culture of I0 P and IE H cells results in cells resistant to both drugs. Cells were plated together, treated with PEG or left untreated, cultured for 20 d, and visualized as in A. (C and D) IE H cells fuse to each other but not to themselves. I0 P and IE H cells were dyed and cultured as indicated for 16 h and visualized by fluorescence microscopy. Heterokaryons (indicated by arrows in C and scored in D) contained both green and blue dyes and at least one green and one blue nucleus. (E) Tissue culture medium from IE H cells induces cell fusion. Tissue culture medium conditioned by either I0 P or IE H cells for 16 h was tested in the fusion assay (see Materials and methods). All experiments were performed at least three times. The data in D and E are from three independent experiments, and the error bars indicate SD.

Article Snippet: Normal human diploid embryonic lung fibroblast cell line IMR90 and normal human diploid skin fibroblast cell lines Detroit 551, HSF43, and Hs68, as well as transformed monkey cell lines COS-1 and CMMT cell lines were obtained from American Type Culture Collection and cultured in 90% DME and 10% FBS and in the absence of antibiotics.

Techniques: Transformation Assay, Cell Culture, Staining, Co-Culture Assay, Fluorescence, Microscopy, Single Vesicle Fusion Assay