human bone marrow stromal cell line hs5  (ATCC)

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Stroma-derived miR-214 coordinates tumor dissemination

doi: 10.1186/s13046-022-02553-5

Figure Lengend Snippet: miR-214 is transferred from stroma to tumor cells. A - D miR-214 and miR-223 expression levels in Extracellular Vesicles (EVs) derived from miR-214 over and miR-214 wt MEFs or from HS5 were previously transduced with a miR-214sponge (miR-214 sponge ) or an empty (control) expressing vector, measured by qRT-PCR analysis. E – F miR-214 expression levels in B16-F10 and MA-2 cells pretreated for 24 h with EVs derived from miR-214 over or miR-214 wt MEFs or from miR-214 sponged HS5 (miR-214 sponge ) or empty controls (control), measured by qRT-PCR analysis. A-F Results are shown as fold changes (mean ± SD of triplicates) relative to controls, normalized on U6. At least 2 independent experiments (with triplicates) were performed and representative results are shown. G-H Expression levels of TFAP2C, ALCAM and ITGA5 in B16-F10 ( G ) and MA-2 ( H ) cells pretreated for 24/48 h with EVs derived from miR-214 over and miR-214 wt MEFs ( G ) or from HS5 ( H ) miR-214sponged HS5 (miR-214 sponge ) or empty controls (control), measured by Western Blot analysis. Protein modulations were calculated relative to controls, normalized on the loading control and expressed as percentages. MEFs = murine embryo fibroblasts; SEM = standard error of mean. SD = standard deviation. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001

Article Snippet: B16-F10 murine melanoma cells, EO771 mouse mammary tumor cells, NIH3T3 fibroblasts and human bone marrow stromal HS5 cell lines were obtained from The American Type Culture Collection.

Techniques: Expressing, Derivative Assay, Transduction, Control, Plasmid Preparation, Quantitative RT-PCR, Western Blot, Standard Deviation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Stroma-derived miR-214 coordinates tumor dissemination

doi: 10.1186/s13046-022-02553-5

Figure Lengend Snippet: Stroma components favor tumor cell motility in a miR-214-dependent manner. A – L Wound healing ( A , B , E , F , G , H , I , J ) or transwell ( C , D , K , L ) assays for mouse B16-F10 or EO771 or human MA-2 or 4175-TGL cells were pretreated for 24 h with conditioned medium (CM) or Extracellular Vesicles (EVs) derived from either miR-214 wt and miR-214 over CAFs ( A , C ) or MEFs ( B , D ) or, from either NIH3T3 mouse fibroblasts ( E - H ) or HS5 human bone marrow stroma cells ( I - L ) previously transduced with a miR-214sponge (miR-214 sponge ) or an empty (control) expressing vector. M Wound healing assay for MA-2 cells pretreated for 24 h with EV-depleted Conditioned Medium (CM) or EVs derived from HS5 cells previously transduced with a miR-214sponge (miR-214 sponge ) or an empty (control) expressing vector. All transwell migration assays were evaluated 18 h later; while wound healing motility assays were evaluated 6 h (B16-F10)—18 h (EO771, MA-2)—24 h (4175-TGL) later. All results are shown as mean ± SEM and respectively shown as mean ± SEM of the area (pixels) or distance/time (μm/h of 10 pictures/duplicates) covered by migrated cells. For ( A , B , E , F , G , H ; I , J , M ) experiments were performed at least twice as duplicates and pooled results of two or three experiments are shown. For ( C , D , K , L ) experiments were performed as triplicates and representative experiments are shown. CAFs = cancer associated fibroblasts; MEFs = murine embryo fibroblasts; SEM = standard error of mean. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001

Article Snippet: B16-F10 murine melanoma cells, EO771 mouse mammary tumor cells, NIH3T3 fibroblasts and human bone marrow stromal HS5 cell lines were obtained from The American Type Culture Collection.

Techniques: Derivative Assay, Transduction, Control, Expressing, Plasmid Preparation, Wound Healing Assay, Migration

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Stroma-derived miR-214 coordinates tumor dissemination

doi: 10.1186/s13046-022-02553-5

Figure Lengend Snippet: Tumor cells influence miR-214 expression in stroma cells. A miR-214 expression levels assessed by qRT-PCR analysis in Stat3 wt or Stat3. ko MEFs, treated with IL-6 for 6 h. B miR-214 expression levels evaluated by qRT-PCR analysis in NIH3T3 or CAFs or HS5 cells following pretreatment with B16-F10 or MA-2 Conditioned Medium (CM) or left untreated (NT) for 6 h. C-D miR-214 expression assessed by qRT-PCR analysis in CAFs treated for 6 h with B16-F10 EV-depleted Conditioned Medium (CM) in presence of 50 μg/ml of anti-IL-6R Ab ( C ) or 10 μg/ml of anti-IL-6 Ab ( D ) or control IgG. E Significant positive correlations between miR-214 expression and different IL-6 and Stat3-related signatures in TCGA-SKCM (top) or TCGA-BRCA (bottom) datasets. Samples are presented in plots and heatmaps (Red = positive; white/blue = negative; grey = no correlation). Non-significant relationships: p -value > 0.05. AZARE_sig ; DAUER_sig ; IL-6_sig [ , ] M5897; ALVAREZ_sig ; TH_sig ; Stat3_sig_up ; Jak/Stat [ , ] M11564; Stat3_sig_down . For ( A - D ) results are presented as fold changes (mean ± SD of triplicates) relative to controls, normalized on U6. At least 2 independent experiments (with triplicates) were performed and either representative results ( A , C ) or pools of all results ( B , D ) are shown. SD = standard deviation; Ab = antibody. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001

Article Snippet: B16-F10 murine melanoma cells, EO771 mouse mammary tumor cells, NIH3T3 fibroblasts and human bone marrow stromal HS5 cell lines were obtained from The American Type Culture Collection.

Techniques: Expressing, Quantitative RT-PCR, Control, Standard Deviation

Journal: PLoS ONE

Article Title: Analysis of acute lymphoblastic leukemia drug sensitivity by changes in impedance via stromal cell adherence

doi: 10.1371/journal.pone.0258140

Figure Lengend Snippet: (A) BLQ5 cells were plated on HS5 stromal cells, resulting in a reduction of impedance (n = 3). (B) Treatment of BLQ5 cells with different concentrations of VX-680 as indicated (n = 3). Dotted lines show the d5 and d9 time points at which cells were harvested for (C) live cell numbers determined by Trypan blue exclusion. Statistical significance was determined using one-way ANOVA, followed by followed by Dunnett’s multiple comparison test. Error bars represent standard deviation. p values relative to DMSO control: ****<0.0001. or (D) Percentage apoptotic cells characterized by FACS using 7-AAD and Annexin V.

Article Snippet: The murine OP9 bone marrow stromal cell line (CRL-2749) and the human HS5 bone marrow stromal cell line (PCS-500-041) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Comparison, Standard Deviation, Control