Journal: Biomolecules & Therapeutics
Article Title: Megakaryocyte-Derived IL-8 Acts as a Paracrine Factor for Prostate Cancer Aggressiveness through CXCR2 Activation and Antagonistic AR Downregulation
doi: 10.4062/biomolther.2023.005
Figure Lengend Snippet: MEG-01-derived IL-8 promotes proliferation and invasion of prostate cancer cells. (A, B) MEG-01 cells were pre-labeled with CMTPX, a red fluorescent cell tracker prior to their placement in 1 μm pore-sized insert. The insert was mounted on the well with cancer cells (LNCaP or PC-3) and incubated in serum-starved (1%) condition for 24 h. The migrated microparticles of MEG-01 cells were visualized using fluorescence microscopy at 200× magnification (A). The number of cancer cells in the lower well was counted (B). The bar graphs represent the means ± SEM from three independent experiments. * p <0.05, compared to LNCaP cells without co-culture. # p <0.05, compared to MEG-01-untreated group in each cell line. (C) Invasion assay of LNCaP and PC-3 in the absence or presence of MEG-01 cells. Cancer cells were seeded in an 8 μm pore-sized insert and mounted on a well that lacked or contained MEG-01 cells, and incubated for 24 h. The invading cancer cells were stained, photographed, and counted (n=3). * p <0.05, compared to LNCaP cell without co-culture. # p <0.05 compared to MEG-01-untreated group in each cell line. (D) Invasion assay of IL-8-silenced PC-3 cells in the absence and presence of MEG-01 cells for 24 h (n=3). * p <0.05, compared to siNT. # p <0.05, compared to si IL-8 -treated group. (E, F) CXCL chemokine levels were measured by ELISA (n=3). CXCL chemokines in culture supernatant of MEG-01 cells without cancer cells (E) and with cancer cell co-culture (F). Culture conditions were the same as described in (A). * p <0.05, compared to MEG-01-untreated group.
Article Snippet: For visualization of diffused MEG-01-derived microparticles, MEG-01 cells were stained with 5 μM of the cell-tracking red fluorescent dye CMTPX (Thermo Fisher Scientific).
Techniques: Derivative Assay, Labeling, Incubation, Fluorescence, Microscopy, Co-Culture Assay, Invasion Assay, Staining, Enzyme-linked Immunosorbent Assay