Journal: Neoplasia (New York, N.Y.)
Article Title: CXCL1-Mediated Interaction of Cancer Cells with Tumor-Associated Macrophages and Cancer-Associated Fibroblasts Promotes Tumor Progression in Human Bladder Cancer
doi: 10.1016/j.neo.2016.08.002
Figure Lengend Snippet: Association of CXCL1 production in TAMs/CAFs with invasion potential of human bladder cancer. (A) Dual immunofluorescence staining analysis for CD204 or αSMA (green) and CXCL1 (red) shows the expression of CXCL1 in TAMs and CAFs in the tumoral area. Overlay images and their magnified images are shown (2 panels on the right). Original magnification, 200×. (B) CXCL1 mRNA expression level was determined by real-time RT-PCR. Expression levels are the result of 3 experiments and are expressed as the mean ± SD relative to that of UROtsa, which is defined as 1. (C) THP-1 and NIH3T3 cells were infected with lentiviral constructs harboring empty vectors (LvNega) or CXCL1 expression vectors (LvCXCL1). Western blot analysis demonstrated altered CXCL1 expression in both cell lines. The changes in the expression levels of CD204 and αSMA were evaluated in infected THP-1 and NIH3T3 cells, respectively. Actin served as the loading control. (D) MGH-U3 and T24 cells were suspended in the conditioned media collected from manipulated TAMs and CAFs and subjected to in vitro invasion assays. Data are expressed as the mean ± SD of 3 independent experiments conducted in triplicate; *, P < 0.05, **, and P < 0.01.
Article Snippet: Four bladder cancer cell lines, J82, UMUC3, T24 (ATCC, Manassas, VA, USA), MGH-U3 (a generous gift from Dr H. LaRue at Laval University Cancer Research Centre, Quebec, Canada), benign bladder cell line UROtsa (a generous gift from Dr Donald Sens at the University of North Dakota School of Medicine, Grand Forks, ND, USA), acute monocytic leukemia cell line THP-1 (ATCC), mouse dermal fibroblast cell line NIH3T3 (ATCC), and HEK293FT (Invitrogen, Carlsbad, CA, USA) were used in the present study.
Techniques: Immunofluorescence, Staining, Expressing, Quantitative RT-PCR, Infection, Construct, Western Blot, In Vitro