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: Virologica Sinica

Article Title: Topoisomerase IIα orchestrates secretion of IL-6 and IL-8 with human papillomavirus replication

doi: 10.1016/j.virs.2025.11.004

Figure Lengend Snippet: Top2α depletion might deregulate inflammation in HPV-positive cells. A RNA-seq analysis was performed using HaCaT31 cells with Top2α knockdown. Differential expressed genes (n = 117) were identified as downstream targets of Top2α. Top20 pathways regulated by Top2α were listed with Metascape online analysis. B RT-qPCR analysis was conducted to assess the expression levels of TOP2A, IL-6, IL-8, IL-1B, IL-18, TNF, IL-17, CCL2, CCL5, CCL20, CXCL9, CXCL10, CXCL11, and IFNB in HaCaT31 cells with Top2α knockdown. GAPDH served as the internal reference gene for normalization. Data are representative of at least three independent biological repeats. The statistical analysis was assayed by 2-tailed t -test. ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, ∗∗∗ P ≤ 0.001, ns = no significance. The red border highlights the mRNA of target genes that were consistently and significantly altered in both shRNA cell lines.

Article Snippet: IL-6 (MCE, #HY-P7044), IL-8 (MCE, #HY-P7379), CCL2 (MCE, #HY-P7237), IL-6 neutralizing antibody (SinoBiological, #10395-MHK23), anti-IL-8 antibody (Selleck, #A2524).

Techniques: RNA Sequencing, Knockdown, Quantitative RT-PCR, Expressing, shRNA

Journal: Virologica Sinica

Article Title: Topoisomerase IIα orchestrates secretion of IL-6 and IL-8 with human papillomavirus replication

doi: 10.1016/j.virs.2025.11.004

Figure Lengend Snippet: Top2α regulates IL-6 and IL-8 secretion in HPV-positive cells. A The protein microarray was utilized to measure the levels of IL-6, IL-8, and CCL2 in vaginal discharge samples from normal, cervical intraepithelial neoplasia (CIN), and cervical cancer patients. B Gene Expression Profiling Interactive Analysis (GEPIA) was performed to analyze the correlation of IL-6, IL-8, and CCL2 expression with the overall survival rate of cervical cancer patients. The red lines represent patients with high gene expression, and the blue lines with low gene expression. C Western blot demonstrating the expression of Top2α, IL-6, IL-8, and CCL2 proteins in the Top2α-depleted cell lines. D The levels of secreted IL-6, IL-8, and CCL2 outside Top2α knockdown cells were determined by ELISA assay. E RT-qPCR was performed to evaluate the expression levels of Top2α, IL-6, and IL-8 in HaCaT cells following Top2α knockdown. F Western blot analysis for the expression of IL-6 and IL-8 in HaCaT cells with Top2α knockdown. GAPDH served as the loading control. G The secretion levels of IL-6, IL-8, and CCL2 were measured by ELISA in HaCaT cells after Top2α knockdown. Data are representative of at least three independent biological repeats. The statistical analysis was assayed by 2-tailed t -test. ∗ P ≤ 0.05, ∗∗∗ P ≤ 0.001, ∗∗∗∗ P ≤ 0.0001. ns = no significance.

Article Snippet: IL-6 (MCE, #HY-P7044), IL-8 (MCE, #HY-P7379), CCL2 (MCE, #HY-P7237), IL-6 neutralizing antibody (SinoBiological, #10395-MHK23), anti-IL-8 antibody (Selleck, #A2524).

Techniques: Microarray, Gene Expression, Expressing, Western Blot, Knockdown, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Control

Journal: Virologica Sinica

Article Title: Topoisomerase IIα orchestrates secretion of IL-6 and IL-8 with human papillomavirus replication

doi: 10.1016/j.virs.2025.11.004

Figure Lengend Snippet: IL-6/IL-8 promotes HPV replication. A ExoV-qPCR was performed to detect HPV episomes in the undifferentiated or differentiated HaCaT31 cells that treated with IL-6 (10 ng/mL) or an anti-IL-6 neutralizing antibody (80 ng/mL) for 72 h. Shown is the relative HPV DNA levels in the treated cells normalized to the untreated cells. B, C Dot blot analysis was applied with the HPV31-specific probe to measure HPV episome levels in the undifferentiated or differentiated HaCaT31 cells that treated with IL-6 (10 ng/mL) or an anti-IL-6 neutralizing antibody (80 ng/mL) for 72 h. Meth-blue served as the loading control. The staining intensities were normalized to the ones of the wild type cells (lower panel). D RT-qPCR analysis of HPV late gene L1 expression in the differentiated cells with IgG, anti-IL-6 or IL-6 treatments as illustrated in figure. E Western blot demonstrating the expression of involucrin and GAPDH proteins. F The extrachromosomal DNA was extracted from undifferentiated or differentiated HaCaT31 cells that treated with IL-8 (10 ng/mL) and an anti-IL-8 neutralizing antibody (50 μg/mL) for 72 h, followed by ExoV-qPCR analysis for E6 levels. G RT-qPCR was used to analyze HPV late gene L1 expression in differentiated cells following treatment with IgG, anti-IL-8, or IL-8. (H) The DNA was extracted from undifferentiated or differentiated HaCaT31 cells that treated with CCL2 (100 ng/mL) for 72 h. Viral episome levels were analyzed by ExoV-qPCR using HPV31 E6 primers. YWHAZ was used as the internal control for data normalization. Data are representative of three biological independent repeats. The statistical analysis was assayed by one-way ANOVA test and means are expressed ±SEM. ∗ P ≤ 0.05, ∗∗ P ≤ 0.01, ∗∗∗ P ≤ 0.001, ∗∗∗∗ P ≤ 0.0001, ns = no significance.

Article Snippet: IL-6 (MCE, #HY-P7044), IL-8 (MCE, #HY-P7379), CCL2 (MCE, #HY-P7237), IL-6 neutralizing antibody (SinoBiological, #10395-MHK23), anti-IL-8 antibody (Selleck, #A2524).

Techniques: Dot Blot, Control, Staining, Quantitative RT-PCR, Expressing, Western Blot

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