Journal: International Journal of Molecular Sciences

Article Title: Ibulocydine Inhibits Migration and Invasion of TNBC Cells via MMP-9 Regulation

doi: 10.3390/ijms25116123

Figure Lengend Snippet: MMP-9 downregulation is critical in IB-induced metastasis inhibition in human TNBC cells. ( a ) After treatment with 1 or 3 µM IB, wound healing scratch assays were performed using TNBC cells. Cell migration was monitored under a phase-contrast microscope for 24 h: Bar, 50 μm. Data are shown as the mean ± SD. * p < 0.05, *** p < 0.001 vs. 24 h untreated control. ( b ) Invasion assays of TNBC cells treated with 0.5 or 1 µM IB for 24 h were performed. Invading cells were stained with crystal violet and observed using a fluorescence microscope (4×). Data are presented as the mean ± SD. *** p < 0.001 vs. untreated control. ( c ) Cell extracts were prepared from cells treated with the indicated IB concentrations for 24 h. MMP-9 and MMP-2 levels were detected using Western blotting. α-tubulin was used as a loading control. ( d ) TNBC cells were treated with the indicated IB concentrations for 24 h. Cell extracts were prepared for Western blotting of mesenchymal markers. ( e ) MMP-9 overexpressing stable cells were scratched during wound healing assays. MMP-9 upregulation was confirmed using Western blotting. Cell migration was monitored for 24 h and observed under a phase-contrast microscope: Bar, 50 μm. The graphs quantitatively show the area of wound recovery. Data are presented as the mean ± SD. *** p < 0.0001 vs. IB-treated vector cells. ( f ) Invasion assays of MMP-9 overexpressing stable cell lines treated with 0.5 or 1 µM IB for 24 h were performed. Invading cells were stained with crystal violet and observed using a fluorescence microscope (4×). Data are shown as the mean ± SD. * p < 0.05, *** p < 0.001 vs. IB-treated vector cells.

Article Snippet: The human TNBC cell line (MDA-MB-231-Luc) was purchased from Caliper Life Sciences (Hopkinton, MA, USA).

Techniques: Inhibition, Migration, Microscopy, Staining, Fluorescence, Western Blot, Plasmid Preparation, Stable Transfection

Journal: Scientific Reports

Article Title: Double-modified, thio and methylene ATP analogue facilitates wound healing in vitro and in vivo

doi: 10.1038/s41598-024-63759-5

Figure Lengend Snippet: Impact of α-thio-β,γ-methylene-ATP on the epithelial–mesenchymal transition- related changes in the model MDA-MB-231 cells. ( A ) Transwell migration rate of cells through 8 μm pore sizes. Data represent the means ± SEM from at least 3 independent experiments, ( B ) Crossing the collagen barrier in the presence of tested compounds. ( C ) The impact of tested analogues on in vivo cells migration in zebrafish xenograft model ( D ) QPCR analysis of EMT markers after treatment with 4a and 4b. Data represent the means ± SEM from at least 3 independent experiments ***p < 0.0005, **p < 0.001, *p < 0.05 compared to the control.

Article Snippet: The MDA-MB-231 (human breast adenocarcinoma) were purchased from Cell Biolabs (San Diego, California, USA, cat no AKR-201).

Techniques: Migration, Analogues, In Vivo

Journal: Scientific Reports

Article Title: Double-modified, thio and methylene ATP analogue facilitates wound healing in vitro and in vivo

doi: 10.1038/s41598-024-63759-5

Figure Lengend Snippet: In vitro verification of ATP analogues activity in the presence of P2Y2 receptor antagonist. ( A ) The impact of tested ATP analogues on Ca 2+ mobilization in HaCaT cells in the presence and absence of P2Y2 antagonist ( B ) The influence of 4a derivative on transwell migration of MDA-MB-231 cells in the presence and absence of P2Y2 antagonist. Data represent the means ± SEM from at least 3 independent experiments. ****p < 0.0001 vs control. C—untreated control cells, RFU—relative fluorescence units. Scale bars 50 μm.

Article Snippet: The MDA-MB-231 (human breast adenocarcinoma) were purchased from Cell Biolabs (San Diego, California, USA, cat no AKR-201).

Techniques: In Vitro, Analogues, Activity Assay, Migration, Fluorescence

Journal: Breast Cancer Research : BCR

Article Title: CXCL17-derived CD11b + Gr-1 + myeloid-derived suppressor cells contribute to lung metastasis of breast cancer through platelet-derived growth factor-BB

doi: 10.1186/s13058-019-1114-3

Figure Lengend Snippet: CXCL17 increased lung metastasis in vivo. a CXCL17 increased the lung metastasis of 4T1 cells in an orthotropic model. The total number of tumor nodule per whole lung lobes was counted and averaged among the animals of each group. b The H&E staining of tumor sections. BALB/c mice were treated with PBS or recombinant mouse CXCL17 protein by intra-tracheal administration for 14 days (1 μg/mouse, 2 times/week, n = 6 per group). 4T1 were implanted into the fat pads of mice. Tumor nodules of 4T1 in the primary site and lungs were collected after 24 days of injections. c CXCL17 increased lung metastasis in human breast MDA-MB-231. Nude mice were treated with PBS or recombinant CXCL17 protein by intra-tracheal administration for 14 days (1 μg/mouse, 2 times/week, n = 6 per group). MDA-MB-231 cells were implanted into mice by tail vein injection. The MDA-MB-231 tumor nodules in the lungs of mice were collected after 90 days of injections. d The H&E staining of tumor sections. e Knockdown of CXCL17 decreased the spontaneous metastatic ability of L4T1 cells. f The H&E staining of tumor sections. CXCL17-knockdown-L4T1 were implanted into the fat pads of mice ( n = 6 per group). Tumor nodules in the lungs were collected after 24 days of injections. Representative lung tissue sections were stained with H&E and photographed at × 100 magnification. Each value is the mean ± SEM; Mann-Whitney U test was performed, * p < 0.05. T tumor

Article Snippet: MDA-MB-231-RFP-Luciferase cells were obtained from GenTarget Inc. All cell lines were assessed for mycoplasma contamination using MycoAlert Mycoplasma Detection Kit (Lonza, Walkersville, MD) every 6 months.

Techniques: In Vivo, Staining, Recombinant, Injection, MANN-WHITNEY

Journal: Breast Cancer Research : BCR

Article Title: CXCL17-derived CD11b + Gr-1 + myeloid-derived suppressor cells contribute to lung metastasis of breast cancer through platelet-derived growth factor-BB

doi: 10.1186/s13058-019-1114-3

Figure Lengend Snippet: CD11b + Gr-1 + myeloid cells in lungs metastatic niche promote cancer cell colonization. a CXCL17 increased cancer cell extravasation into the lungs of mice. Athymic nude mice were treated with recombinant CXCL17 protein (1 μg/mouse, 2 times/week, n = 6 per group) for 14 days; MDA-MB-231-RFP-Luc cells were injected into mice by tail vein. The lungs of these mice were harvested after 48-h injection, then examined by a confocal microscope. b Scheme of CD11b + Gr-1 + depletion in the animal model. Depletion of CD11b + Gr-1 + cells decreased CXCL17-mediated cancer extravasation ( c ) and tumor nodules formation ( d ) in the lungs of mice. e The H&E staining of tumor sections of lungs. For MDSC depletion studies, mice ( n = 6 per group) were treated with isotype or anti-Gr-1 antibodies (Bio X Cell) at 100 μg/mouse intraperitoneal injections every 4th day, and 4T1 cells were injected via tail vein into the mice ( n = 6 per group). The control group received intraperitoneal injection of purified rat immunoglobulins. PKH26-labeled 4T1 cells were injected into mice by tail vein for indicated times (24 days for lung metastasis and 48 h for extravasation). The lungs of these mice were harvested and examined by a confocal microscope. Each value is the mean ± SEM; * p < 0.05. White arrows indicate cancer cells

Article Snippet: MDA-MB-231-RFP-Luciferase cells were obtained from GenTarget Inc. All cell lines were assessed for mycoplasma contamination using MycoAlert Mycoplasma Detection Kit (Lonza, Walkersville, MD) every 6 months.

Techniques: Recombinant, Injection, Microscopy, Animal Model, Staining, Purification, Labeling