Journal: Pharmaceutics

Article Title: CD44 Targeting of Cisplatin-Loaded Hyaluronic Acid-Modified Mesoporous Silica Nanoparticles for Lung Adenocarcinoma: Synthesis, Characterization, In Vitro and In Vivo Evaluation

doi: 10.3390/pharmaceutics18020171

Figure Lengend Snippet: In vitro drug releases of Cisplatin from CDDP, MSN-CDDP and HA-MSN-CDDP in phosphate-buffered saline (PBS): ( a ) pH 7.4, ( b ) pH 5.5. ( c ) Fluorescence microscope image of cellular uptake of control group (scale bar 10 µm) ( d ) and Rhodamine B-labeled HA-MSN by A549-Luc-C8 cells (scale bar 10 µm). ( e ) PBMC, ( f ) CCD-34Lu, ( g ) A549-Luc-C8 cell viabilities after 48 h of treatment to 1, 2, 10, 20, 100 and 200 µg/mL (according to Cisplatin concentration) of CDDP, MSN, HA-MSN, MSN-CDDP and HA-MSN-CDDP. ( h ) Apoptosis rate after CDDP, MSN-CDDP and HA-MSN-CDDP treatment (according to IC 50 concentrations on A549-Luc-C8 cells). Data is presented as mean± SD from three repeats and three independent experiments. * Statistically significant difference compared to the control group ( p < 0.05).

Article Snippet: A549-Luc-C8 (Luciferase-expressing human non-small cell lung carcinoma, CVCL-5J13) and CCD-34Lu (Human lung fibroblast cells, ATCC-CRL-1491TM) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) Ham’s F12.

Techniques: In Vitro, Saline, Fluorescence, Microscopy, Control, Labeling, Concentration Assay

Journal: Molecular Cell

Article Title: The TRAIL-Induced Cancer Secretome Promotes a Tumor-Supportive Immune Microenvironment via CCR2

doi: 10.1016/j.molcel.2017.01.021

Figure Lengend Snippet: TRAIL Induces a Cytokine Secretome (A) Schematic of the quantitative, label-free secretome profiling approach. (B and C) (B) A pool of all secreted proteins that were upregulated by isoleucine zipper (iz)-TRAIL (100 ng/mL) (>2-fold induction as cutoff) was analyzed by label-free quantification and searched for (C) the most significantly enriched GO molecular functions. Line thickness is representative of the associated p value. Chemokine activity, p = 0.0005; chemokine receptor binding, p = 0.0006; cytokine activity, p = 0.0039. (D) A549 cells were stimulated with iz-TRAIL (100 ng/mL). After 24 hr, cytokine concentrations in the cell supernatants were measured via ELISA; cell viability was determined by CellTiter-Glo. p values were obtained by two-tailed Student’s t test. ∗ p ≤ 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Data are presented as mean ± SEM, n = 3. See also Figure S1 .

Article Snippet: A549-luc , Caliper Life Science , Bioware Cell Line A549-Luc-C8.

Techniques: Activity Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Molecular Cell

Article Title: The TRAIL-Induced Cancer Secretome Promotes a Tumor-Supportive Immune Microenvironment via CCR2

doi: 10.1016/j.molcel.2017.01.021

Figure Lengend Snippet: Surviving Cancer Cells Produce Cytokines upon TRAIL Stimulation (A) A549, HCT116 WT, and HCT116 BAX/BAK KO cells were stimulated with the indicated concentrations of iz-TRAIL for 24 hr; cell viability was determined by CellTiter-Glo. (B) HCT116 WT or HCT116-BAX/BAK KO cells were stimulated with iz-TRAIL (100 ng/mL) for 24 hr; cell viability was determined by CellTiter-Glo and cytokine concentrations in the cell supernatants were measured by ELISA. (C) HCT116 WT cells were pre-incubated with QVD (10 μM) or DMSO for 30 min, followed by addition of iz-TRAIL (100 ng/mL for 24 hr; cell viability and cytokine concentrations were determined as above. (D) A549 cells were pre-incubated with PIK-75 (100 nM) or QVD (10 μM) + PIK-75 (100 nM) for 30 min, followed by addition of iz-TRAIL (100 ng/mL); cell viability and IL-8 concentration were determined as above. Data are presented as mean ± SEM, n = 3. See also Figure S2 .

Article Snippet: A549-luc , Caliper Life Science , Bioware Cell Line A549-Luc-C8.

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Concentration Assay

Journal: Molecular Cell

Article Title: The TRAIL-Induced Cancer Secretome Promotes a Tumor-Supportive Immune Microenvironment via CCR2

doi: 10.1016/j.molcel.2017.01.021

Figure Lengend Snippet: Caspase-8 and FADD Presence Is Required for TRAIL-Mediated Cytokine Induction (A) A549 WT cells were transiently transfected with small interfering RNA (siRNA) against the indicated targets for 48 hr and stimulated with QVD (10 μM) (CTRL) or QVD (10 μM) + iz-TRAIL (100 ng/mL); 24 hr later, cytokine concentrations were determined by ELISA. A representative western blot is shown. (B) A549 WT or FADD KO, either empty vector (+EV) or FADD (+FADD) reconstituted, were stimulated with iz-TRAIL (100 ng/mL) for 24 hr, followed by determination of cytokine concentrations by ELISA. (C) Cells as in (B) were stimulated with iz-TRAIL (100 ng/mL) for the indicated times, followed by immunoblotting for the indicated proteins. Data are presented as mean ± SEM, n ≥ 3. A representative experiment of n = 2 is shown. See also and .

Article Snippet: A549-luc , Caliper Life Science , Bioware Cell Line A549-Luc-C8.

Techniques: Transfection, Small Interfering RNA, Enzyme-linked Immunosorbent Assay, Western Blot, Plasmid Preparation

Journal: Molecular Cell

Article Title: The TRAIL-Induced Cancer Secretome Promotes a Tumor-Supportive Immune Microenvironment via CCR2

doi: 10.1016/j.molcel.2017.01.021

Figure Lengend Snippet: FADD Promotes Tumor Growth and Accumulation of Alternatively Activated Myeloid Cells (A) Severe combined immunodeficiency (SCID)/beige mice were injected with 2 × 10 6 A549 WT or FADD KO cells stably expressing luciferase into the lateral tail vein. Tumor burden was assessed after 24 days via bioluminescence imaging. n = 11/group. Representative images are shown. (B) Histological quantification of tumor burden. Representative images of H&E-stained lung sections (5× magnification) are shown. (C) C57BL/6 mice were injected with 5 × 10 5 3LL cells into the lateral tail vein. Lung weights were determined 28 days later. Representative lungs are shown. (D) Histological quantification of tumor burden in lungs from mice shown in (C). Representative images of H&E-stained lung sections (5× magnification) are shown. (E) The indicated cytokines were quantified in lung homogenates by ELISA. (F and G) Absolute number of (F) CD11b + Gr1 + or (G) CD11b + Gr1 + CD206 + cells within tumor-bearing lungs. Unpaired, two-tailed Student’s t test was performed to determine significance. ∗ p ≤ 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Data are represented as mean ± SEM. See also Figure S5 .

Article Snippet: A549-luc , Caliper Life Science , Bioware Cell Line A549-Luc-C8.

Techniques: Injection, Stable Transfection, Expressing, Luciferase, Imaging, Staining, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Molecular Cell

Article Title: The TRAIL-Induced Cancer Secretome Promotes a Tumor-Supportive Immune Microenvironment via CCR2

doi: 10.1016/j.molcel.2017.01.021

Figure Lengend Snippet: A TRAIL-Induced Secretome Polarizes Human Monocytes to Alternatively Activated Myeloid Cells (A) Schematic of the monocyte polarization protocol. (B–D) CD14 + cells were isolated from healthy donor peripheral blood mononuclear cells (PBMCs) via magnetic CD14 + microbeads and incubated with untreated or iz-TRAIL-treated (100 ng/mL) WT or FADD KO A549 conditioned medium (CM). After 48 hr, treated myeloid cells were stained with fluorochrome-labeled antibodies against HLA-DR, CD14, and CD206 and analyzed by flow cytometry. Data are presented as fold change in HLA-DR lo/neg CD14 + cells (B), HLA-DR lo/neg , CD14 + , CD206 + cells (C), or HLA-DR + , CD14 + CD206 + macrophages (D) upon iz-TRAIL-induced CM incubation. Unpaired, two-tailed Student’s t test was performed to determine significance. ns = p > 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Data are presented as mean ± SEM, n = 4.

Article Snippet: A549-luc , Caliper Life Science , Bioware Cell Line A549-Luc-C8.

Techniques: Isolation, Incubation, Staining, Labeling, Flow Cytometry, Two Tailed Test

Journal: Molecular Cell

Article Title: The TRAIL-Induced Cancer Secretome Promotes a Tumor-Supportive Immune Microenvironment via CCR2

doi: 10.1016/j.molcel.2017.01.021

Figure Lengend Snippet:

Article Snippet: A549-luc , Caliper Life Science , Bioware Cell Line A549-Luc-C8.

Techniques: Western Blot, Flow Cytometry, Recombinant, In Vivo, Lysis, Blocking Assay, Enzyme-linked Immunosorbent Assay, Proliferation Assay, Sequencing, Plasmid Preparation, shRNA, Software