Journal: Cancer Research

Article Title: MEN1 Promotes Ferroptosis by Disrupting CD44 Alternative Splicing to Suppress Lung Cancer

doi: 10.1158/0008-5472.CAN-25-0021

Figure Lengend Snippet: MEN1 -modulated ferroptosis is required for suppressing lung tumor growth and metastasis. A–D, Cell viability and representative invasion and migration images of WT and KO NCI-H460 cells; invaded cells and migration rate were quantified at the indicated time points. E, Schematic of experimental lung metastasis model in M-NSG mice. F, Representative IHC (Ki67 and HMGB1) and H&E staining images of lung tissue sections from M-NSG mice xenografted with WT/KO NCI-H460 cells. Scale bars, 1 mm or 50 μm. G and H, Tumor burden ( G ) and quantification of Ki67/HMGB1 IHC staining in lung tissues ( H ) from xenografted M-NSG mice. I, Schematic of the primary mouse lung cancer study ( n = 7, 9, and 8 for WT, KS, and KMS cohorts). J–M, Representative lung pictures ( J ), H&E staining and Ki67/HMGB1 IHC images of lung tissues ( K ), tumor burden and tumor number ( L ), and quantification of Ki67/HMGB1 IHC staining ( M ). Scale bars, 1 mm or 50 μm. Black arrows, tumor nodules. N, Kaplan–Meier survival analysis of WT, KS, and KMS mice. O, Lipid peroxidation (LPO) levels in lungs from WT, KS, and KMS mice. a.u, arbitrary unit. P and Q, Representative RNA FISH images ( P ) and quantification of GPX4 probe numbers in WT lung tissues and KS/KMS lung tumor tissues ( Q ). Scale bars, 20 or 50 μm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant.

Article Snippet: The paraffin-embedded tissue sections after deparaffinization and rehydration were subjected to antigen retrieval in 0.1 mol/L citrate buffer (pH 6.0) in a steam pot for 15 minutes, blocked with 5% BSA for 1 hour, and incubated with primary antibodies including menin (Bethyl Laboratories, #A300-105A, RRID: AB_2143306), Ki67 (Cell Signaling Technology, #9129, RRID: AB_2687446), HMGB1 (Proteintech, #10829-1-AP, RRID: AB_2232989), and SFTPC (Abclonal, #A1835, RRID: AB_2763872).

Techniques: Migration, Staining, Immunohistochemistry

Journal: Signal Transduction and Targeted Therapy

Article Title: An RNA–RNA crosstalk network involving HMGB1 and RICTOR facilitates hepatocellular carcinoma tumorigenesis by promoting glutamine metabolism and impedes immunotherapy by PD-L1+ exosomes activity

doi: 10.1038/s41392-021-00801-2

Figure Lengend Snippet: a Prediction of candidate HMGB1 crosstalk RNAs using the ceRDB database ( https://www.oncomir.umn.edu/cefinder ). The top 12 candidate RNAs and scores were shown. b The RNA expression levels of HMGB1 and the 12 candidate RNAs were analyzed in early stage HCC (BCLC stages 0 and A HCC) tissues by qRT-PCR. HBV + HCC (above) n = 26; HBV − HCC (below), n = 9. The results are means ± SD. Names marked red and black represent significantly ( p < 0.05) and nonsignificant genes, respectively. c Pearson correlation coefficient analysis between expression levels of HMGB1 and the 12 candidate RNAs, respectively, in early stage HCC tumor tissues

Article Snippet: The frozen sections were examined based on in situ hybridization using digoxin-labeled HMGB1, RICTOR, and miR-429 probes and digoxin detection kit (Boster biological technology, Guangzhou, China) according to the suggested procedures.

Techniques: RNA Expression, Quantitative RT-PCR, Expressing

Journal: Signal Transduction and Targeted Therapy

Article Title: An RNA–RNA crosstalk network involving HMGB1 and RICTOR facilitates hepatocellular carcinoma tumorigenesis by promoting glutamine metabolism and impedes immunotherapy by PD-L1+ exosomes activity

doi: 10.1038/s41392-021-00801-2

Figure Lengend Snippet: a The mRNA expression of HMGB1 and RICTOR in the 11 indicated cell lines as determined by qRT-PCR. * p < 0.05. b Pearson correlation coefficient analysis between the expression level of HMGB1 and RICTOR in the cell lines from ( a ). c Immunohistochemical staining of HMGB1 and RICTOR in liver tissues in the DEN + CCl4-induced liver cancer mouse model. d , e The expression levels of RICTOR and HMGB1 mRNA (left) and protein (right) after the interference of HMGB1 or RICTOR in the HCCLM3 ( d ) and PLC/PRF/5 ( e ) cell lines (abbreviated as “KD-HMGB1 or KD-RICTOR”, respectively). f The expression levels of RICTOR and HMGB1 mRNA (left) and protein (right) after overexpression of HMGB1 or RICTOR 3′UTR in the QSG-7701 cell line (abbreviated as “OE-HMGB1 or OE-RICTOR”, respectively). The mRNA levels (left) were determined by qRT-PCR and protein level (right) was determined by Western blot. * p < 0.05, *** p < 0.001

Article Snippet: The frozen sections were examined based on in situ hybridization using digoxin-labeled HMGB1, RICTOR, and miR-429 probes and digoxin detection kit (Boster biological technology, Guangzhou, China) according to the suggested procedures.

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemical staining, Staining, Over Expression, Western Blot

Journal: Signal Transduction and Targeted Therapy

Article Title: An RNA–RNA crosstalk network involving HMGB1 and RICTOR facilitates hepatocellular carcinoma tumorigenesis by promoting glutamine metabolism and impedes immunotherapy by PD-L1+ exosomes activity

doi: 10.1038/s41392-021-00801-2

Figure Lengend Snippet: a Luciferase assays with reporter constructs containing the wild-type or mutant HMGB1/RICTOR 3′UTR downstream of the luciferase gene were performed after co-transfection with miR-200a/200b/429 mimics in HEK293T cells. * p < 0.05, ** p < 0.01, *** p < 0.001. b The mRNA levels of HMGB1 and RICTOR were determined by qRT-PCR 48 h after transfection of 100 nM miR-200a/200b/429 mimics in indicated cells. ** p < 0.01, *** p < 0.001. c RNA FISH assays detecting the cellular localization of HMGB1/RICTOR and miR-429 in indicated cells

Article Snippet: The frozen sections were examined based on in situ hybridization using digoxin-labeled HMGB1, RICTOR, and miR-429 probes and digoxin detection kit (Boster biological technology, Guangzhou, China) according to the suggested procedures.

Techniques: Luciferase, Construct, Mutagenesis, Cotransfection, Quantitative RT-PCR, Transfection

Journal: Signal Transduction and Targeted Therapy

Article Title: An RNA–RNA crosstalk network involving HMGB1 and RICTOR facilitates hepatocellular carcinoma tumorigenesis by promoting glutamine metabolism and impedes immunotherapy by PD-L1+ exosomes activity

doi: 10.1038/s41392-021-00801-2

Figure Lengend Snippet: a Determination of spheroid formation after 3000 cells were seeded in low-adhesion plates for 10 days (left). The number of tumor spheroids was quantitated (right). *** p < 0.001. b Cell proliferation was evaluated using CCK8 assays. The results are means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001. c Male nude mice ( n = 5) were subcutaneously injected with 1 × 10 4 HMGB1/RICTOR 3′UTR over-expressed liver cells (abbreviated as “OE-HMGB1 or OE-RICTOR”, respectively) or HMGB1/RICTOR mRNA interference HCC cells (abbreviated as “KD-HMGB1 or KD-RICTOR”, respectively). Tumorigenesis was assessed after 48 days as shown

Article Snippet: The frozen sections were examined based on in situ hybridization using digoxin-labeled HMGB1, RICTOR, and miR-429 probes and digoxin detection kit (Boster biological technology, Guangzhou, China) according to the suggested procedures.

Techniques: Injection

Journal: Signal Transduction and Targeted Therapy

Article Title: An RNA–RNA crosstalk network involving HMGB1 and RICTOR facilitates hepatocellular carcinoma tumorigenesis by promoting glutamine metabolism and impedes immunotherapy by PD-L1+ exosomes activity

doi: 10.1038/s41392-021-00801-2

Figure Lengend Snippet: a PMBC killing assay scheme. HMGB1/RICTOR mRNA interference HCC cells (abbreviated as “KD-HMGB1 or KD-RICTOR”, respectively) were cocultured with activated PBMCs and treated with or without Atezolizumab for 48 h before apoptosis detection. b Cell apoptosis in treated HCC cells was evaluated by TUNEL assay (above). The apoptotic cell ratios were shown (below). c Exosomes isolated from cell culture supernatants in treated HCC cells were determined by flow cytometry using PE-fluorescein-conjugated anti-PD-L1 antibody (red). Isotype matched antibody (PE-fluorescein-conjugated anti-IgG antibody) was used as gating controls (black). Percentages of PD-L1 + Exo resulted from the relative fluorescence values gated by isotype controls. Exo: exosomes. ** p < 0.01, *** p < 0.001

Article Snippet: The frozen sections were examined based on in situ hybridization using digoxin-labeled HMGB1, RICTOR, and miR-429 probes and digoxin detection kit (Boster biological technology, Guangzhou, China) according to the suggested procedures.

Techniques: TUNEL Assay, Isolation, Cell Culture, Flow Cytometry, Fluorescence

Journal: Signal Transduction and Targeted Therapy

Article Title: An RNA–RNA crosstalk network involving HMGB1 and RICTOR facilitates hepatocellular carcinoma tumorigenesis by promoting glutamine metabolism and impedes immunotherapy by PD-L1+ exosomes activity

doi: 10.1038/s41392-021-00801-2

Figure Lengend Snippet: a A model of the RNA–RNA crosstalk network involving HMGB1 and RICTOR acting in the early stage HCC

Article Snippet: The frozen sections were examined based on in situ hybridization using digoxin-labeled HMGB1, RICTOR, and miR-429 probes and digoxin detection kit (Boster biological technology, Guangzhou, China) according to the suggested procedures.

Techniques:

Journal: Nature Communications

Article Title: Cell surface patching via CXCR4-targeted nanothreads for cancer metastasis inhibition

doi: 10.1038/s41467-024-47111-z

Figure Lengend Snippet: In stage I, Nanothread-1 recognizes and multivalently binds CXCR4, aligning adjacent receptors while presenting coiled motif decoys on the cancer cell surface. Subsequently in stage II, Nanothread-2 engages these decoys, intertwining with Nanothread-1 into a coiled-coil supramolecular network. This sequential actuation is proposed to generate physical dragging forces that cluster an expanded vicinity of receptors, synchronizing CXCR4 clustering and amplifying mechanotransduction to enable adequate manipulation of downstream signaling events. Concurrently, photosensitizers ‘hitchhike’ on Nanothread-2 to the tumor site for targeted photodynamic therapy (PDT), inducing immunogenic cell death (ICD) to transform the primary tumor into an in-situ vaccine. Nanothread ‘patching’ on CXCR4 superclusters is proposed to seal spontaneous metastatic potential by disrupting the metastasis cascade — reshaping metastatic tumor cell ‘seeds’, intercepting ‘seed-soil’ crosstalk, and regressing the pre-metastatic niche ‘soil’. Moreover, manipulated CXCR4 clustering downstream effects, including survival pathway interference, hypoxia alleviation and immunosuppression reversal, are expected to sensitize the tumor to PDT. Consequently, a localized anti-tumor immune response would be initiated against the primary tumor, while also establishing an abscopal memory effect against disseminated metastases. CRT, calreticulin; ATP, adenosine triphosphate; HMGB1 high mobility group protein B1; IFN-γ, interferon-γ; MDSC, myeloid-derived suppressor cells; EMT, epithelial–mesenchymal transition; TDSF, tumor-derived secreted factors; PMN, pre-metastatic niche; CXCL12, Chemokine (C-X-C Motif) Ligand 12. Created with BioRender.com.

Article Snippet: QuantiCyto® Human/Mouse/Rat HMGB1 ELISA Kit (Cat No.: EHRC01.96) was provided by Neobioscience Technology Co,Ltd.

Techniques: In Situ, Derivative Assay

Journal: Nature Communications

Article Title: Cell surface patching via CXCR4-targeted nanothreads for cancer metastasis inhibition

doi: 10.1038/s41467-024-47111-z

Figure Lengend Snippet: a , b Primary tumor growth, survival, and lung metastasis in spontaneous lung metastasis mouse models of female BALB/c mice orthotopically bearing murine 4T1 breast tumors. Timelines indicate schedules for four cycle treatments with P-PS, P-BS-PS or P-BS-CM1 → P-PS-CM2 without ( a ) or with ( b ) laser irradiation. Metastatic nodules were indicated with red circles. Scale bar was 400 μm for photographs of whole lungs (top) and was 100 μm for hematoxylin-eosin staining images (bottom). c , d Intratumoral level of calreticulin (CRT) exposure, adenosine triphosphate (ATP) secretion, and high mobility group protein B1 (HMGB1) release in primary tumor tissues on Day 28 after three cycle treatments without ( c ) and with ( d ) laser irradiation. e Representative immunofluorescent images of interferon-γ (IFN-γ) and quantification of IFN-γ + CD8 + T lymphocytes in primary tumors on Day 28 after three cycle treatments with laser irradiation. Blue indicates cell nuclei. Red indicates IFN-γ. Scale bars, 50 μm. f Tumor growth and survival rate over time after cotreatment with P-BS-CCM1 → P-PS-CCM2 (+) and CD8-depleting antibodies. n = 8 animals per group for primary tumor growth and survival studies in ( a , b , f ). n = 5 animals per group for other studies. Data are presented as mean ± SD. Statistics for survival curves in ( a , b , f ) are calculated by log-rank (Mantel–Cox) test. Statistics of others are calculated by one-way ANOVA with Tukey’s multiple comparisons without adjustments. NS not significant, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. Source data are provided as a Source Data file.

Article Snippet: QuantiCyto® Human/Mouse/Rat HMGB1 ELISA Kit (Cat No.: EHRC01.96) was provided by Neobioscience Technology Co,Ltd.

Techniques: Irradiation, Staining