Journal: European Journal of Medical Research

Article Title: PEAK1 promotes prostate cancer progression and docetaxel resistance by mediating the polarization of tumor-associated macrophages

doi: 10.1186/s40001-025-03568-2

Figure Lengend Snippet: Association of PEAK1 level and “M2” polarization of TAM in PCa. A , B Level of immune cell infiltration between the high and low PEAK1 expression groups. C–H Gene correlation analysis between PEAK1 and IL-10, TGF- β , CCL2, Arg1, CD163, and CD206 in PRAD was conducted via GEPIA ( http://gepia.cancer-pku.cn/ ). I IHC was conducted to detect CCL2 in the tumor tissues. Scale bar = 50 μm. J , K Western blotting was conducted to measure CCL2, Arg1, CD163, and CD206 in the tumor tissues. L IHC was conducted to detect CD45, iNOS, CD163, CD8, PD-L1, granzyme B, and IFN-γ in the tumor tissues. Scale bar = 50 μm. N = 3. * stands for p < 0.05, ** stands for p < 0.01, *** stands for p < 0.001

Article Snippet: Next, the levels of CCL2 and IL-6 in the cell supernatant were determined via CCL2 (#EK187EG) and IL-6 (#EK106) kits (Multi Sciences, Hangzhou, China), respectively.

Techniques: Expressing, Western Blot

Journal: European Journal of Medical Research

Article Title: PEAK1 promotes prostate cancer progression and docetaxel resistance by mediating the polarization of tumor-associated macrophages

doi: 10.1186/s40001-025-03568-2

Figure Lengend Snippet: Functions of PCa cells with PEAK1 upregulation on “M2” polarization of macrophages. A Co-culture system graph. THP1 cells were put in the lower chamber and PC3 or DU145 cells were put in the upper chamber. B RT-PCR was conducted to detect CCL2 and IL-6 mRNA expression in PC3 and DU145 cells. C , D ELISA was conducted to test the levels of CCL2 and IL-6 in the co-culture medium. E RT-PCR was conducted to detect the expressions of “M2” markers, including IL-10, TGF- β , CCL2, Arg1, CD163, and CD206 in THP1 cells co-cultured with PEAK1-overexpressed PCa cells. F Migration of THP1 cells was evaluated using a transwell assay. G , H RT-PCR and WB analysis of CCR2 expression in THP1 cells co-cultured with PEAK1-overexpressed or normal PC3/DU145 cells. I To test the functions of “M2” macrophages on the PCa cells, the THP1 cells were put in the upper transwell chamber, while the PC3 cells were put in the lower chamber. J CCK8 assay was performed to detect the viability of PC3 cells. K EdU-staining assay was performed to evaluate the proliferation of PC3 cells. The EdU-positive cell rate was counted. L Transwell assay was carried out to evaluate cell migration. M WB analysis for detecting E -cad, Vim, Snail, and Twist1. N CCK-8 assay was carried out to determine the IC₅₀ value of DTX in PC3 cells. O Colony formation assays were performed to determine the colony-forming ability of cells with DTX treatment. P WB analysis for assessing the expressions of Bcl-2, Bax and cleaved caspase-3 (c-Casp3). N = 3. Q , R PC3 cells and DU145 cells were, respectively, treated with TGF- β (10 ng/mL) or LY2157299 (10 μM), a selective TGF- β receptor type I (TGF–βRI) kinase inhibitor. RT-PCR and WB were conducted to detect the alteration of CCR2 expression in the two cells. * stands for p < 0.05, ** stands for p < 0.01, *** stands for p < 0.001

Article Snippet: Next, the levels of CCL2 and IL-6 in the cell supernatant were determined via CCL2 (#EK187EG) and IL-6 (#EK106) kits (Multi Sciences, Hangzhou, China), respectively.

Techniques: Co-Culture Assay, Reverse Transcription Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Migration, Transwell Assay, CCK-8 Assay, Staining

Journal: European Journal of Medical Research

Article Title: PEAK1 promotes prostate cancer progression and docetaxel resistance by mediating the polarization of tumor-associated macrophages

doi: 10.1186/s40001-025-03568-2

Figure Lengend Snippet: Mechanistic diagram of PEAK1 in PCa. Summary diagram of the functions and mechanisms of PEAK1 in PCa. Upregulated PEAK1 promotes the HIF-1α/STAT3/NF-κB pathway activation, thus enhancing tumor proliferation, migration, EMT, and reducing apoptosis, docetaxel/enzalutamide sensitivity. PEAK1 enhances CCL2, IL-6 and PD-L1 expression from PCa cells, which mediate “M2” polarization of TAMs and inducing TGF- β expression from TAMs. TGF- β can act on PCa cells and induce PEAK1 upregulation

Article Snippet: Next, the levels of CCL2 and IL-6 in the cell supernatant were determined via CCL2 (#EK187EG) and IL-6 (#EK106) kits (Multi Sciences, Hangzhou, China), respectively.

Techniques: Activation Assay, Migration, Expressing