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

Journal: Cell Communication and Signaling : CCS

Article Title: Fusobacterium nucleatum -reprogrammed adipocytes promote tumor cisplatin resistance through the CCL2-CCR2 axis in the necrotic metastatic neck nodes of head and neck carcinoma

doi: 10.1186/s12964-025-02550-z

Figure Lengend Snippet: F. nucleatum -reprogrammed adipocytes facilitate tumor cisplatin-resistance through activation of CREB/HSL and secretion of CCL2. A Western blot analysis of phosphorylation level of CREB/HSL in adipocytes and F. nucleatum -cocultured adipocytes ( n = 3). B Western blot analysis of lipophagy-related genes in adipocytes and F. nucleatum -cocultured adipocytes ( n = 3). C PCA plot showed the overall differences in transcriptional profile of adipocytes and F. nucleatum -cocultured adipocytes. D The circle heatmap of differential genes in adipocytes and F. nucleatum -cocultured adipocytes. E KEGG pathway enrichment. F MA plot of differential genes with cytokine-related genes highlighted. G ELISA analysis of CCL2 content in the supernatant of adipocytes and F. nucleatum -cocultured adipocytes ( n = 3). H Changes in CCL2 expression in inguinal adipose tissue following F. nucleatum infection. AdiCM: adipocyte supernatant. FN: Fusobacterium nucleatum . Adi: adipocytes. Data are presented as means ± SEM. * P < 0.05,‌** P < 0.01, ‌*** P < 0.001, **** P < 0.0001, ns: no statistical significance

Article Snippet: The ELISA quantitation of CCL2 was performed using Mouse MCP-1/CCL2 ELISA Kit (Boster Biological Technology, China).

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Enzyme-linked Immunosorbent Assay, Expressing, Infection

Journal: Biology Direct

Article Title: The role of SH3RF2 in lung squamous cell carcinoma and M2 polarization: insights into LZTS2 ubiquitination

doi: 10.1186/s13062-025-00677-0

Figure Lengend Snippet: ( A ) Cell viability of SKMES1 cells treated with ICG-001. ( B ) RNA level of CCL2 in SKMES1 cells. ( C ) Protein level of CCL2 in SKMES1 cells. ( D ) Cellular migration and invasion ability of THP-1 cells analyzed by transwell assay. ( E ) RNA levels of Arg-1, CD163 and IL-10 in M0 THP-1 cells treated with SKMES1 CM. ( F ) The proportion of CD206 + cells in M0 THP-1 cells with SKMES1 CM treatment. EV: empty vector, SR oe : SH3RF3 overexpression. **: p < 0.01, ***: p < 0.001

Article Snippet: Colonies were fixed with 4% paraformaldehyde at room temperature for 25 min and stained with crystal violet dye for 5 min. CCL2 levels were determined using the Human CCL2 ELISA Kit (Lianke Bio, China).

Techniques: Migration, Transwell Assay, Plasmid Preparation, Over Expression

Journal: Biology Direct

Article Title: The role of SH3RF2 in lung squamous cell carcinoma and M2 polarization: insights into LZTS2 ubiquitination

doi: 10.1186/s13062-025-00677-0

Figure Lengend Snippet: ( A & B ) SKMES1 cells with SH3RF2 overexpression or knockdown were injected subcutaneously into the right armpit of Balb/c nude mice. Tumor size was measured every 3 days. Tumors were removed and weighed after 33 days. ( C & D ) The protein level of CCL2 in tumor determined by ELISA. ( E ) Detection of SH3RF2 expression in tumor tissues by immunohistochemistry, bar = 50 μm. ( F ) Immunohistochemistry for Ki-67 in tumor tissues, bar = 50 μm. ( G ) The proportion of CD11b + F4/80 + CD206 + cells in tumor. NC sh : negative control shRNA, SR sh−1 : SH3RF2 shRNA-1, SR sh−2 : SH3RF2 shRNA-2, EV: empty vector, SR oe : SH3RF3 overexpression. **: p < 0.01, ***: p < 0.001

Article Snippet: Colonies were fixed with 4% paraformaldehyde at room temperature for 25 min and stained with crystal violet dye for 5 min. CCL2 levels were determined using the Human CCL2 ELISA Kit (Lianke Bio, China).

Techniques: Over Expression, Knockdown, Injection, Enzyme-linked Immunosorbent Assay, Expressing, Immunohistochemistry, Negative Control, shRNA, Plasmid Preparation

Journal: Biology Direct

Article Title: The role of SH3RF2 in lung squamous cell carcinoma and M2 polarization: insights into LZTS2 ubiquitination

doi: 10.1186/s13062-025-00677-0

Figure Lengend Snippet: ( A ) Protein level of LZTS2 in SKMES1 cells with SH3RF2 and LZTS2 overexpression. ( B ) Cell viability of SKMES1 cells with SH3RF2 and LZTS2 overexpression. ( C ) Immunofluorescence detection of the distribution of β-catenin in SKMES1 cells and quantitative analysis of nuclear-to-cytoplasmic ratio of β-catenin fluorescence intensity. ( D ) Protein level of CCL2 in SKMES1 cells. ( E ) RNA levels of Arg-1, CD163 and IL-10 in M0 THP-1 cells with SKMES1 CM treatment. ( F ) Cellular migration and invasion ability of M0 THP-1 cells with SKMES1 CM treatment. ( G ) The proportion of CD206 + cells in the M0 THP-1 cells treated with SKMES1 CM. EV: empty vector, SR oe : SH3RF3 overexpression. LZ oe : LZTS2 overexpression. **: p < 0.01, ***: p < 0.001

Article Snippet: Colonies were fixed with 4% paraformaldehyde at room temperature for 25 min and stained with crystal violet dye for 5 min. CCL2 levels were determined using the Human CCL2 ELISA Kit (Lianke Bio, China).

Techniques: Over Expression, Immunofluorescence, Fluorescence, Migration, Plasmid Preparation