Journal: International Journal of Medical Sciences

Article Title: Discovering the abnormalities and functional importance of ferroptosis-related molecules in cervical cancer

doi: 10.7150/ijms.107133

Figure Lengend Snippet: Search for key genes. (A) The random survival forest algorithm was used to screen the characteristic genes and rank the importance of 10 prognosis-related genes. (B) LASSO regression identified 25 genes as characteristic genes. (C) Venn diagram showing the number of genes common to and unique to the random survival forest analysis and LASSO regression. (D-F) Survival analysis of the key genes, including SQLE, GCH1 and H1.2. (G) Verification of SQLE, GCH1 and H1.2 from GEO. The expression levels of GCH1 and H1.2 were increased significantly in the tumor groups. (H-I) Differential expression of GCH1 and H1.2 in the external validation sets GSE7410 (H) and GSE7803 (I) in the tumor and normal groups.

Article Snippet: Subsequently, primary stainings for GCH1 (1:200, 28501-1-AP, Proteintech), CD11c (45581T, CST, 1:3000, used as an M1 macrophage marker), and CD163 (ab182422, Abcam, 1:5000, used as an M2 macrophage marker) were performed in the same manner.

Techniques: Expressing, Quantitative Proteomics, Biomarker Discovery

Journal: International Journal of Medical Sciences

Article Title: Discovering the abnormalities and functional importance of ferroptosis-related molecules in cervical cancer

doi: 10.7150/ijms.107133

Figure Lengend Snippet: Relationships between the key genes and immune infiltration. (A) The immune cell contents of the normal and tumor groups. (B) Immune cell correlation map. (C) Comparison of immune cells between the tumor and normal groups. (D, E) Correlations between CIBERSORT and the expression levels of H1.2 and GCH1.

Article Snippet: Subsequently, primary stainings for GCH1 (1:200, 28501-1-AP, Proteintech), CD11c (45581T, CST, 1:3000, used as an M1 macrophage marker), and CD163 (ab182422, Abcam, 1:5000, used as an M2 macrophage marker) were performed in the same manner.

Techniques: Comparison, Expressing

Journal: International Journal of Medical Sciences

Article Title: Discovering the abnormalities and functional importance of ferroptosis-related molecules in cervical cancer

doi: 10.7150/ijms.107133

Figure Lengend Snippet: GSVA analysis of the key genes. (A) Enriched signaling pathways associated with highly expressed GCH1. (B) Enriched signaling pathways associated with highly expressed H1.2.

Article Snippet: Subsequently, primary stainings for GCH1 (1:200, 28501-1-AP, Proteintech), CD11c (45581T, CST, 1:3000, used as an M1 macrophage marker), and CD163 (ab182422, Abcam, 1:5000, used as an M2 macrophage marker) were performed in the same manner.

Techniques: Protein-Protein interactions

Journal: International Journal of Medical Sciences

Article Title: Discovering the abnormalities and functional importance of ferroptosis-related molecules in cervical cancer

doi: 10.7150/ijms.107133

Figure Lengend Snippet: GSEA of GCH1 (A) and H1.2 (B).

Article Snippet: Subsequently, primary stainings for GCH1 (1:200, 28501-1-AP, Proteintech), CD11c (45581T, CST, 1:3000, used as an M1 macrophage marker), and CD163 (ab182422, Abcam, 1:5000, used as an M2 macrophage marker) were performed in the same manner.

Techniques:

Journal: International Journal of Medical Sciences

Article Title: Discovering the abnormalities and functional importance of ferroptosis-related molecules in cervical cancer

doi: 10.7150/ijms.107133

Figure Lengend Snippet: Immunohistochemical verification of GCH1 and H1.2. (A) Localization of GCH1 and H1.2 in cervical cancer tissues (×40 and ×200). (B) Expression levels of GCH1 and H1.2 in cervical cancer tissue and paracarcinoma tissue. (C) OS/PFS of patients with cervical cancer with high GCH1/H1.2 expression compared with those with low GCH1/H1.2 expression.

Article Snippet: Subsequently, primary stainings for GCH1 (1:200, 28501-1-AP, Proteintech), CD11c (45581T, CST, 1:3000, used as an M1 macrophage marker), and CD163 (ab182422, Abcam, 1:5000, used as an M2 macrophage marker) were performed in the same manner.

Techniques: Immunohistochemical staining, Expressing

Journal: International Journal of Medical Sciences

Article Title: Discovering the abnormalities and functional importance of ferroptosis-related molecules in cervical cancer

doi: 10.7150/ijms.107133

Figure Lengend Snippet: Immunofluorescence spatial distance analysis of GCH1. (A and B) Expression and localization of GCH1, CD11c, CD163 and CD8 in cervical cancer tissues. (C) Total cell numbers in Low GCH1 and High GCH1 cervical cancer tissues. (D-F) CD11c+ M1, CD163+ M2 and CD8+ T immune cell proportions in Low GCH1 and High GCH1 cervical cancer tissues. (G) M1/M2 cell ratios in Low GCH1 and High GCH1 cervical cancer tissues. (H-J) Distances among CD11c+ M1, CD163+ M2 and CD8+ T immune cells and tumor cells in Low GCH1 and High GCH1 cervical cancer tissues.

Article Snippet: Subsequently, primary stainings for GCH1 (1:200, 28501-1-AP, Proteintech), CD11c (45581T, CST, 1:3000, used as an M1 macrophage marker), and CD163 (ab182422, Abcam, 1:5000, used as an M2 macrophage marker) were performed in the same manner.

Techniques: Immunofluorescence, Expressing

Journal: International Journal of Medical Sciences

Article Title: Discovering the abnormalities and functional importance of ferroptosis-related molecules in cervical cancer

doi: 10.7150/ijms.107133

Figure Lengend Snippet: Relationship between GCH1 and the biological behavior of cervical cancer. (A) Relative mRNA expression levels of GCH1 in cervical cancer cell lines. (B) Relative mRNA expression levels of GCH1 in SiHa and CaSki cells transfected with si-GCH. (C-D) Cell cycle distribution of SiHa cells transfected with si-GCH1. (E-F) Apoptosis of SiHa cells transfected with si-GCH1. (G-H) Migration and invasion abilities of SiHa cells transfected with si-GCH1. (I) Viability of GCH1-knockdown SiHa cells, as determined by CCK-8 assay. (J) Expression levels of p-PI3K, p-AKT, p-mTOR, and GCH1 in SiHa knockdown cells. β-actin was used as an endogenous control. Each experiment was repeated at least 3 times. The data are presented as the means ± SEMs, *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001.

Article Snippet: Subsequently, primary stainings for GCH1 (1:200, 28501-1-AP, Proteintech), CD11c (45581T, CST, 1:3000, used as an M1 macrophage marker), and CD163 (ab182422, Abcam, 1:5000, used as an M2 macrophage marker) were performed in the same manner.

Techniques: Expressing, Transfection, Migration, Knockdown, CCK-8 Assay, Control

Journal: International Journal of Medical Sciences

Article Title: Discovering the abnormalities and functional importance of ferroptosis-related molecules in cervical cancer

doi: 10.7150/ijms.107133

Figure Lengend Snippet: Effects of GCH1 in cancer cells on macrophage polarization. (A) Morphology of THP-1/M0 macrophages in the GCH1-knockdown and control groups. (B) Relative mRNA expression levels of M1 and M2 marker genes.

Article Snippet: Subsequently, primary stainings for GCH1 (1:200, 28501-1-AP, Proteintech), CD11c (45581T, CST, 1:3000, used as an M1 macrophage marker), and CD163 (ab182422, Abcam, 1:5000, used as an M2 macrophage marker) were performed in the same manner.

Techniques: Knockdown, Control, Expressing, Marker

Journal: Cell Reports Medicine

Article Title: ENPP1 inhibitor with ultralong drug-target residence time as an innate immune checkpoint blockade cancer therapy

doi: 10.1016/j.xcrm.2025.102336

Figure Lengend Snippet: STF-1623 boosts intratumoral cGAMP levels to suppress murine breast cancer growth and metastasis (A–C) cGAMP levels (A), IFN-γ levels (B), and relative Ifn-γ mRNA expression (C) in subcutaneous EMT6 (top) or EMT6 shEnpp1 (bottom) tumors at time points indicated after STF-1623 (50 mg/kg) injection. For cGAMP measurement, background measured in vehicle controls was subtracted from the measurements. Mean ± SEM is plotted, n = 3 for EMT6 tumors; n = 4–5 for EMT6 shEnpp1 tumors. (D) BALB/c female mice with established subcutaneous EMT6 tumors received treatment as indicated. Mean ± SEM is plotted. (E) Percent lung metastatic burden determined by hematoxylin and eosin staining of mice in (D) on day 29 of the study. Mean ± SD is plotted, n = 9–10 mice. (F) Immunohistochemistry of CD8 + T cells in randomly selected tumors of mice in (D) on day 29 of the study. Mean ± SD is plotted, n = 5 mice. (G) BALB/c female mice with established orthotopic 4T1 tumors received treatment as indicated. Mean ± SEM of tumor volume is plotted. (H) BALB/c female mice with established orthotopic 4T1 tumors received treatment as indicated. Mice were sacrificed when exhibiting symptoms related to metastasis. The p value for the Kaplan-Meier analysis was determined by log rank test. p values were determined by two-sided unpaired t test unless otherwise mentioned. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; p value is shown if between 0.05 and 0.1. See also .

Article Snippet: To characterize tumor infiltrating lymphocytes, CD8 primary antibody staining (CST, #98941; 1:400) followed by anti-rabbit poly-HRP-IgG (Leica, #DS9800) of tumor samples were performed.

Techniques: Expressing, Injection, Staining, Immunohistochemistry

Journal: Cell Reports Medicine

Article Title: ENPP1 inhibitor with ultralong drug-target residence time as an innate immune checkpoint blockade cancer therapy

doi: 10.1016/j.xcrm.2025.102336

Figure Lengend Snippet: STF-1623 delays murine colorectal tumor growth (A) C57BL/6 female mice with established subcutaneous MC38 tumors received treatment as indicated. Mean ± SEM of tumor volume is plotted. (B) Percent weight change of mice in (A) on day 14 compared to day 1 of the study. Mean ± SD is plotted, n = 10 mice per group. (C) ENPP1 activity of WT, ENPP1 WT-OE , and ENPP1 T238A-OE MC38 cell lysates assessed by [ 32 P] cGAMP hydrolysis by thin-layer chromatography. (D) MC38 ENPP1 WT-OE and MC38 ENPP1 T238A-OE cells were subcutaneously injected in WT female BALB/c mice. n = 9 for ENPP1 WT-OE and n = 10 for ENPP1 T238A-OE . Mean ± SEM of tumor volume is plotted. (E) C57BL/6 female mice with established subcutaneous MC38 ENPP1 WT-OE tumors received treatment as indicated. Mean ± SEM of tumor volume is plotted, n = 8–9 mice. (F) Percent weight change of mice in (E) on day 14 of study compared to day 1 of the study. Mean ± SD is plotted, n = 8 or 9 mice per group. (G) BALB/c mice with established CT26 tumors received treatment as indicated. Mean ± SEM is plotted, n = 10 mice. (H) Percent weight change of mice in (G) on day 13 compared to day 1 of the study. Mean ± SD is plotted, n = 10 mice. (I) BALB/c mice with established subcutaneous CT26 tumors received 5 or 50 mg/kg of STF-1632 on days 9–11, 16–18 post inoculation ( pi ), 10 mg/kg anti-PD-1 intraperitoneally on day 9, 12, 16, and 19 pi , or a combination of the two. On day 23 of the study, tumors were isolated and processed for flow cytometry. The number of CD3 + T cells, CD4 + CD3 + T cells, and CD8 + CD3 + T cells per gram of tumors is shown. Mean ± SEM is plotted, n = 10 mice. p values were calculated by two-sided unpaired t test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: To characterize tumor infiltrating lymphocytes, CD8 primary antibody staining (CST, #98941; 1:400) followed by anti-rabbit poly-HRP-IgG (Leica, #DS9800) of tumor samples were performed.

Techniques: Activity Assay, Thin Layer Chromatography, Injection, Isolation, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: ENPP1 inhibitor with ultralong drug-target residence time as an innate immune checkpoint blockade cancer therapy

doi: 10.1016/j.xcrm.2025.102336

Figure Lengend Snippet: STF-1623 controls Panc02 pancreatic tumor growth by activating anti-cancer immunity (A and B) BALB/c female mice with established subcutaneous Panc02 tumors received treatment indicated. On day 12 of the study, tumors were processed for flow cytometry: the percentage of IA-IE + CD206 low M1 macrophage (F4/80 + GR-1 − ) over IA-IE − CD206 high M2 macrophage, the number of CD335 + CD3 − NK cells, CD335 + CD3 + NKT cells, CD4 + CD3 + T cells, and CD8 + CD3 + T cells per gram of tumors, and the ratio of CD8 + T cells over Foxp3 + CD4 + Treg cells (A), CD69 + CD4 + T cells, PD-1 + CD4 + T cells, Ki67 + CD4 + T cells, CD69 + CD8 + T cells, PD-1 + CD8 + T cells, and Ki67 + CD8 + T cells (B). Mean ± SD is plotted, n = 6 mice for vehicle and STF-1623 groups, and n = 3 for all other combination therapy groups. (C) BALB/c female mice with established Panc02 tumors received the treatment as indicated. Mean ± SEM of tumor volume is plotted, n = 9–12 mice. Spider plots of individual tumor growth are shown. (D) Percent weight change of mice in (C) on day 45 compared to day 1 of the study. Mean ± SD is plotted, n = 9–12 mice. NK, natural killer; NKT, natural killer T; Treg, regulatory T cells; IR. ionizing radiation. p values were calculated by two-sided unpaired t test unless otherwise noted. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001; p value is shown if between 0.05 and 0.1.

Article Snippet: To characterize tumor infiltrating lymphocytes, CD8 primary antibody staining (CST, #98941; 1:400) followed by anti-rabbit poly-HRP-IgG (Leica, #DS9800) of tumor samples were performed.

Techniques: Flow Cytometry