Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of IL-10, TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Sections were incubated with primary antibodies against Ki-67 (Servicebio, Cat# GB111499 ), E-cadherin (Proteintech, Cat# 20874-1-AP), and Vimentin (Proteintech, Cat# 10366-1-AP).

Techniques: Knockdown, Functional Assay, Quantitative RT-PCR, Flow Cytometry, Marker, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Derivative Assay, Co-Culture Assay, Western Blot, Migration

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 depletion in macrophages restrains LUAD growth and metastasis in vivo. (A) Schematic of orthotopic lung implantation and experimental metastasis models using LLC cells mixed with BMDMs expressing shNC or sh-circSMAD4. (B) Representative images of orthotopic lung tumors. (C) Tumor weight of orthotopic implants. (D) Overall survival of mice bearing orthotopic tumors. (E) Immunofluorescence showing F4/80 and circSMAD4 signals in tumor tissues. Scale bar, 50 μm. (F, G) Representative Ki-67 IHC staining and quantification in orthotopic tumors. Scale bar, 50 μm. (H) Representative bioluminescence images of lung tumor burden in the metastasis model. (I) Tumor weight in the metastasis model. (J) Overall survival of mice in the metastasis model. (K–M) Representative IHC staining and quantification of E-cadherin and vimentin in tumors. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Sections were incubated with primary antibodies against Ki-67 (Servicebio, Cat# GB111499 ), E-cadherin (Proteintech, Cat# 20874-1-AP), and Vimentin (Proteintech, Cat# 10366-1-AP).

Techniques: In Vivo, Expressing, Immunofluorescence, Immunohistochemistry

Journal: Genes & Diseases

Article Title: Identification of PLXNC1 as a novel biomarker for consensus molecular subtype 4 in colorectal cancer

doi: 10.1016/j.gendis.2025.101974

Figure Lengend Snippet: PLXNC1 knockdown inhibits gene signatures characteristic of CMS4 colorectal cancer. (A) qRT-PCR analysis was applied to examine the relative mRNA expression of genes related to epithelial–mesenchymal transition (EMT), complement, angiogenesis, and immunosuppression. GAPDH was a normalization control. (B) Protein expression and quantification results of SNAIL, Vimentin, and E-cadherin in colorectal cancer cells were determined by western blotting. (C) Immunofluorescence analysis of the levels of E-cadherin (the epithelial marker, red) and Vimentin (the mesenchymal marker, green) proteins in the control and PLXNC1-silenced LoVo cells. The nuclei were stained with Hoechst (blue). Scale bars = 75 μm ∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05.

Article Snippet: Fixed cells were incubated with primary antibody against E-cadherin (1:50; sc-8426; Santa Cruz Biotechnology, California, USA) or vimentin (1:100; BM0135; Boster, Wuhan, China) at 4 °C overnight, followed by incubation with secondary antibodies (1:200; Boster, Wuhan, China) at room temperature for 1 h. The nuclei of cells were stained with Hoechst for 10 min.

Techniques: Knockdown, Quantitative RT-PCR, Expressing, Control, Western Blot, Immunofluorescence, Marker, Staining

Journal: Genes & Diseases

Article Title: Identification of PLXNC1 as a novel biomarker for consensus molecular subtype 4 in colorectal cancer

doi: 10.1016/j.gendis.2025.101974

Figure Lengend Snippet: PLXNC1 promotes tumor growth and metastasis in vivo . (A) Magnetic resonance imaging (MRI) assessment of wild-type (WT) mice carrying subcutaneous tumors. (B) Tumor volumes calculated by MRI. The tumor volume = area of tumor in each slice × slice thickness. (C) Representative images of tumors from WT mice after subcutaneous inoculation of SL4 cells infected with the control vector or sh-PLXNC1 vector. (D) Mice were killed 14 days following subcutaneous injection, and tumors were dissected and weighed. (E) Bioluminescence images showing tumor metastasis by tracking luciferase-expressing SL4 cells in sh-NC and sh-PLXNC1 groups. (F) Histogram showing the bioluminescent signal intensity analyzed by the IVIS System. (G) MRI inspection of hepatic tumor metastasis of colon cancer after intrasplenic injection of SL4 cells in sh-NC and sh-PLXNC1 groups. (H) Gross examination of hepatic tumor metastasis of colon cancer after intrasplenic injection of SL4 cells in sh-NC and sh-PLXNC1 groups. (I) Mice were sacrificed on day 10 after intrasplenic injection, and livers were excised and weighed. (J) Immunohistochemical staining was used to detect the expression levels of PCNA, CD31, TGF-β, vimentin, β-catenin, and E-Cadherin in tumor tissues from sh-NC and sh-PLXNC1 groups. Scale bars = 50 μm. (K) Quantification of immunohistochemical staining in subcutaneous tumor tissues from sh-NC and sh-PLXNC1 groups. (L) Immunohistochemical staining showing expression of epithelial–mesenchymal transition (EMT) markers in liver metastasis tumor tissues from sh-NC and sh-PLXNC1 groups. (M) Quantification of immunohistochemical staining in liver metastasis tumor tissues from sh-NC and sh-PLXNC1 groups. ∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05; ns, not significant.

Article Snippet: Fixed cells were incubated with primary antibody against E-cadherin (1:50; sc-8426; Santa Cruz Biotechnology, California, USA) or vimentin (1:100; BM0135; Boster, Wuhan, China) at 4 °C overnight, followed by incubation with secondary antibodies (1:200; Boster, Wuhan, China) at room temperature for 1 h. The nuclei of cells were stained with Hoechst for 10 min.

Techniques: In Vivo, Magnetic Resonance Imaging, Infection, Control, Plasmid Preparation, Injection, Luciferase, Expressing, Immunohistochemical staining, Staining