Journal: Frontiers in immunology

Article Title: Loss of NLRP3 reduces oxidative stress and polarizes intratumor macrophages to attenuate immune attack on endometrial cancer.

doi: 10.3389/fimmu.2023.1165602

Figure Lengend Snippet: FIGURE 1 NLRP3 levels are significantly reduced in macrophages from EMC compared to NEM. The expression of NLRP3 was examined in macrophages from EMC and NEM, using a GEO database GSE117970. (A) PCA plot. (B) A volcano map to show significant downregulation of NLRP3 in EMC- macrophages. (C) Pathway analysis of differentially expressed genes by Metascape. (D, E) Array reads for NLRP3 (D) and IL-1b (E) in macrophages from EMC and NEM. *p <0.05.

Article Snippet: EMC cell line and modulation of -rimary macrophages A human EMC cell line (HEC-1A, a subclone of HEC-1B (HTB-113, ATCC)) that was isolated from endometrial adenocarcinoma of a 71-year-old patient (19).

Techniques: Expressing

Journal: Frontiers in immunology

Article Title: Loss of NLRP3 reduces oxidative stress and polarizes intratumor macrophages to attenuate immune attack on endometrial cancer.

doi: 10.3389/fimmu.2023.1165602

Figure Lengend Snippet: FIGURE 3 NLRP3-depletion in macrophages increases the growth, invasion, and migration potential of co-cultured EMC cells. BMDMs were isolated from Lys2-Cre; NLRP3(fx/fx) mice as M0, which were further differentiated into M1 macrophages by treatment with IPS and IFNg or differentiated into M2 macrophages by treatment with IL-4 and IL-13. (A–E) M0, M1, or M2 macrophages were co-cultured with HEC-1A. (A) CCK-8 assay for growth of HEC-1A cells. (B, C) Invasion of HEC-1A cells, shown by quantification (B) and by representative images (C). (D, E) Migration potential of HEC-1A cells, shown by quantification (D) and by representative images (E). *p <0.05. N = 5. Scale bars are 100 µm.

Article Snippet: EMC cell line and modulation of -rimary macrophages A human EMC cell line (HEC-1A, a subclone of HEC-1B (HTB-113, ATCC)) that was isolated from endometrial adenocarcinoma of a 71-year-old patient (19).

Techniques: Migration, Cell Culture, Isolation, CCK-8 Assay

Journal: Frontiers in immunology

Article Title: Loss of NLRP3 reduces oxidative stress and polarizes intratumor macrophages to attenuate immune attack on endometrial cancer.

doi: 10.3389/fimmu.2023.1165602

Figure Lengend Snippet: FIGURE 5 NLRP3 depletion promotes growth and metastasis of EMC in vivo. Luciferase-transduced HEC-1A cells were subcutaneously transplanted into mice with macrophage-depletion of NLRP3, Lys2-Cre; NLRP3(fx/fx) and control NLRP3(fx/fx). The tumor formation and metastasis were tested after 90 days. (A, B) Bioluminescence in a luciferin assay, shown by representative images (A) and by quantification (B). (C) Incidence of lung metastasis. (D) Flow cytometry analysis for cytotoxic CD3+CD8+ T cells in tumors. Arrows pointed to lung tissue, and arrowheads pointed to a metastatic tumor in the lung. *p <0.05. N = 10 in each group. Scale bars are 100 µm.

Article Snippet: EMC cell line and modulation of -rimary macrophages A human EMC cell line (HEC-1A, a subclone of HEC-1B (HTB-113, ATCC)) that was isolated from endometrial adenocarcinoma of a 71-year-old patient (19).

Techniques: In Vivo, Luciferase, Control, Flow Cytometry

Journal: Frontiers in immunology

Article Title: Loss of NLRP3 reduces oxidative stress and polarizes intratumor macrophages to attenuate immune attack on endometrial cancer.

doi: 10.3389/fimmu.2023.1165602

Figure Lengend Snippet: FIGURE 6 NLRP3-depletion-induced M2 macrophage polarization reduces cytotoxic T cells in vivo. The tumor was digested and subjected to flow cytometry analysis. (A, B) FACS for CD68 and CD163, shown by quantification (A) and by representative flow charts (B). (C, D) FACS for CD3 and CD8, shown by quantification (C) and by representative flow charts (D). (E) Schematic of the study, showing that loss of NLRP3 polarizes intratumor macrophages to attenuate the immune attack on EMC both directly and indirectly through regulating cytotoxic T cells. *p <0.05. N = 5.

Article Snippet: EMC cell line and modulation of -rimary macrophages A human EMC cell line (HEC-1A, a subclone of HEC-1B (HTB-113, ATCC)) that was isolated from endometrial adenocarcinoma of a 71-year-old patient (19).

Techniques: In Vivo, Cytometry

Journal: Scientific Reports

Article Title: Integrative network toxicology and experimental evidence reveal mechanisms underlying diethyl phthalate-induced initiation and progression of endometrial cancer

doi: 10.1038/s41598-026-39325-6

Figure Lengend Snippet: DEP inhibited proliferation and induced ROS accumulation in EC cells. ( A , B ) CCK-8 assay showing the effect of different concentrations of DEP on proliferation of HEC-1-A ( A ) and Ishikawa ( B ) cells at 0, 24, and 48 h (n = 3). ( C , D ) Representative EdU staining images of HEC-1-A ( C ) and Ishikawa ( D ) cells treated with DEP at the indicated concentrations (DMSO, DEP at 10 nM, 1 μM, and 10 μM). Nuclei were counterstained with Hoechst 33,342 (blue), and EdU-positive cells are shown in red. Scale bar = 100 μm. ( E ) Quantification of EdU-positive cells in HEC-1-A and Ishikawa cell lines (n = 3). ( F ) Representative fluorescence images of ROS staining in HEC-1-A and Ishikawa cells following DEP treatment. White arrows indicate cells with increased ROS levels, and blue arrows indicate cells with lower ROS levels (n = 3). Scale bar = 20 μm. Data are presented as mean ± SD; statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test. (* P < 0.05, *** P < 0.001, **** P < 0.0001).

Article Snippet: The human EC cell lines HEC-1-A and Ishikawa were obtained from GeneChem Gene Technology Co., Ltd. (Shanghai, China).

Techniques: CCK-8 Assay, Staining, Fluorescence

Journal: Scientific Reports

Article Title: Integrative network toxicology and experimental evidence reveal mechanisms underlying diethyl phthalate-induced initiation and progression of endometrial cancer

doi: 10.1038/s41598-026-39325-6

Figure Lengend Snippet: DEP induced cell cycle arrest and modulated the PI3K/AKT signaling pathway in EC cells. ( A ) Flow cytometry analysis of cell cycle distribution in Ishikawa and HEC-1-A cells treated with DEP at the indicated concentrations (DMSO, DEP at 10 nM, 1 μM, and 10 μM). ( B ) Quantification of the proportion of cells in G0/G1, S, and G2/M phases (n = 3). ( C ) WB analysis of PI3K, P-PI3K, AKT, P-AKT, ERK, P-ERK, CDK4, Cyclin D1, GSK3β, and P-GSK3β expression in Ishikawa and HEC-1-A cells after DEP treatment. ( D ) Densitometric quantification of protein expression levels normalized to GAPDH or β-actin (n = 3). Data are presented as mean ± SD; statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test. (ns = not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: The human EC cell lines HEC-1-A and Ishikawa were obtained from GeneChem Gene Technology Co., Ltd. (Shanghai, China).

Techniques: Flow Cytometry, Expressing

Journal: Molecular Medicine Reports

Article Title: MicroRNA-195 inhibits epithelial-mesenchymal transition by targeting G protein-coupled estrogen receptor 1 in endometrial carcinoma

doi: 10.3892/mmr.2019.10652

Figure Lengend Snippet: Overexpression of miR-195 exerts an inhibitory effect on AN3-CA and Hec1A cell viability. To examine the effects of miR-195 on EC development, the miR-195 mimics vector was transfected into human EC (AN3-CA and Hec1A) cells. (A and C) The transfection efficiency was determined by RT-qPCR. (B and D) Cell viabilities at 12, 24 and 48 h of each experimental group were detected using the CCK-8 kit. Each value represents the mean ± SEM (n=3). U6 was considered as an internal control. a P<0.05, aa P<0.01 vs. Control group; b P<0.05, bb P<0.01 vs. Mock group. EC, endometrial carcinoma.

Article Snippet: Human endometrial cancer cell lines (AN3-CA and Hec1A) were obtained from the American Type Culture Collection (ATCC).

Techniques: Over Expression, Plasmid Preparation, Transfection, Quantitative RT-PCR, CCK-8 Assay, Control