Journal: Cell Reports Medicine

Article Title: LTA4H improves the tumor microenvironment and prevents HCC progression via targeting the HNRNPA1/LTBP1/TGF-β axis

doi: 10.1016/j.xcrm.2025.102000

Figure Lengend Snippet:

Article Snippet: The next day, the cells were treated with DEN (25 ng/mL), LTB4 (100 nM, Cayman, 20110) and inhibitors including JNK-IN-8 (2.5 μM, Aladdin, J125321), LY255283 (5 μM, MCE, HY-15744) and U-75302 (200 nM, Cayman, 70705), as indicated in the figures.

Techniques: Microarray, Recombinant, Lysis, Protease Inhibitor, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Reverse Transcription, Activity Assay, Immunoprecipitation, Extraction, Purification, In Vitro, Transfection, Sequencing, Mass Cytometry, ChIP-sequencing, Transgenic Assay, Software

Journal: Military Medical Research

Article Title: Elevated FBXL6 activates both wild-type KRAS and mutant KRAS G12D and drives HCC tumorigenesis via the ERK/mTOR/PRELID2/ROS axis in mice.

doi: 10.1186/s40779-023-00501-8

Figure Lengend Snippet: Fig. 2 FBXL6 facilitates KRASG12D-driven hepatocarcinogenesis and lung metastasis. LSL-Fbxl6KI/+;Alb-Cre (LC), LSL-KrasG12D/+;Alb-Cre (KC) and LSL -KrasG12D/+;LSL-Fbxl6KI/+;Alb-Cre (KLC) mice were monitored for 320 d and then sacrificed. a Representative images of tumorigenesis in LC, KC and KLC mice. b Quantification of the tumor number, tumor size, and liver/body weight ratio of LC (n = 13), KC (n = 10), and KLC mice (n = 12). qPCR was utilized to measure the expression of HCC markers (Cd44, Afp, Gpc3 and Ly6d) (c), proliferation markers (Ki67, Pcna, Ccnb1, and Ccnb2) (d), and metastasis-related markers (Icam1, Vcam1, Mmp9 and Ccl2) (e), in WT liver tissues, LC tumors, KC tumors and KLC tumors. Lung tissues were collected from LC, KC and KLC mice for HE and IHC staining. Representative images of HE and IHC staining for lipase C (LIPC) showing distinct lung metastatic foci expressing LIPC (f). Scale bars = 200 or 50 μm. The ratio of lung metastasis in each cohort was calculated (g). Data are represented as the mean ± SEM. One-way ANOVA was used in (b–e). ns non‑significant; *P < 0.05; **P < 0.01; ***P < 0.001. Fbxl6 F-box and leucine-rich repeat 6, Kras kirsten rat sarcoma, Afp alpha fetoprotein, Gpc3 glypican 3, Ly6d lymphocyte antigen 6 family member D, Ki67 marker of proliferation Ki-67, Pcna proliferating cell nuclear antigen, Ccnb1 cyclin B1, Ccnb2 cyclin B2, Icam1 intercellular adhesion molecule 1, Vcam1 vascular cell adhesion molecule 1, Mmp9 matrix metallopeptidase 9, Ccl2 C–C motif chemokine ligand 2, IHC immunohistochemistry, SEM standard error of the mean

Article Snippet: Mouse strain generation and mouse breeding Mice containing Loxp-STOP-Loxp-Fbxl6 (LSL-Fbxl6KI/+, n = 5) were generated by Biocytogen Pharmaceuticals (Beijing, China), and floxed alleles of Fbxl6 (conditional knockout mice of Fbxl6, CKO-Fbxl6KO/+, n = 4) were generated by Cyagen Biosciences Corporation (Guangzhou, China).

Techniques: Expressing, Immunohistochemistry, Marker

Journal: Military Medical Research

Article Title: Elevated FBXL6 activates both wild-type KRAS and mutant KRAS G12D and drives HCC tumorigenesis via the ERK/mTOR/PRELID2/ROS axis in mice.

doi: 10.1186/s40779-023-00501-8

Figure Lengend Snippet: Fig. 5 mTOR and MEK inhibition significantly blocks hepatocarcinogenesis and lung metastasis triggered by Fbxl6 elevation and Kras mutation. a Nude mice bearing KLC orthotopic HCC tumors were randomized into three groups: the negative control (NC) group, everolimus (E) group and everolimus combined with trametinib (E + T) group. The NC group was treated with vehicle, the E group was treated with everolimus (diluted in DMSO and coil oil, 4 mg/kg, intraperitoneal injection) twice per week for six rounds, and the E + T group was treated with both everolimus (the everolimus dose was as in the E group) and trametinib (diluted in DMSO and coil oil, 0.5 mg/kg, intraperitoneal injection) twice per week for six rounds. The mice were sacrificed after the last injection, and representative images showing tumorigenesis are presented. n = 7. b Tumor weight, the tumor/liver weight ratio, and tumor volume were analyzed (n = 7). c Representative images showing the effect of mTOR and MEK/ERK inhibitors on lung metastases. The lung metastasis rate (d) and number of lung distant lung metastatic foci (e) were quantified. f Representative HE and IHC staining images for lipase C (LIPC) showing distinct lung metastatic foci expressing LIPC. Scale bars = 200 or 50 μm. g Representative images of HE and IHC staining for Prelid2, p-mTOR, p-4EBP1, and p-ERK in the NC, E and E + T groups. Representative consecutive IHC staining images are presented. Scale bars = 200 or 50 μm. One-way ANOVA with Tukey’s multiple comparisons test was used in (b, e). *P < 0.05; **P < 0.01; ***P < 0.001. KLC LSL-KrasG12D/+;LSL-Fbxl6KI/+;Alb-Cre, mTOR mammalian target of rapamycin, MEK mitogen-activated protein kinase kinase, Fbxl6 F-box and leucine-rich repeat 6, Kras kirsten rat sarcoma, NC negative control, E everolimus, T trametinib, DMSO dimethyl sulfoxide, ERK extracellular signal-regulated kinase, IHC immunohistochemistry

Article Snippet: Mouse strain generation and mouse breeding Mice containing Loxp-STOP-Loxp-Fbxl6 (LSL-Fbxl6KI/+, n = 5) were generated by Biocytogen Pharmaceuticals (Beijing, China), and floxed alleles of Fbxl6 (conditional knockout mice of Fbxl6, CKO-Fbxl6KO/+, n = 4) were generated by Cyagen Biosciences Corporation (Guangzhou, China).

Techniques: Inhibition, Mutagenesis, Negative Control, Injection, Immunohistochemistry, Expressing

Journal: Military Medical Research

Article Title: Elevated FBXL6 activates both wild-type KRAS and mutant KRAS G12D and drives HCC tumorigenesis via the ERK/mTOR/PRELID2/ROS axis in mice.

doi: 10.1186/s40779-023-00501-8

Figure Lengend Snippet: Fig. 6 Elevated PRELID2 is positively correlated with the FBXL6/p-ERK/p-mTOR pathway and poor prognosis of HCC. a Representative images of IHC staining for FBXL6, PRELID2, p-ERK, and p-mTOR in human HCC tumors. Scale bar = 50 µm. b The association between PRELID2 and FBXL6, p-ERK, or p-mTOR in 129 paired HCC tumors and adjacent normal tissues was analyzed by χ2 test. c The prognostic significance of PRELID2 in HCC patients was evaluated by Kaplan–Meier analysis. High expression of PRELID2 predicted a shorter overall survival (OS) time. d The association between FBXL6 and p-ERK/p-mTOR protein levels in 129 HCC tissues was evaluated by the χ2 test. e The prognostic significance of the coexpression of FBXL6 and p-ERK in HCC patients was evaluated by Kaplan–Meier analysis. f Kaplan–Meier survival curves showing the overall survival of FBXL6high/ p-ERKhigh HCC patients with high or low PRELID2 expression. PRELID2 the proteins of relevant evolutionary and lymphoid interest (PRELI) domain 2, FBXL6 F-box and leucine-rich repeat 6, ERK extracellular signal-regulated kinase, mTOR mammalian target of rapamycin, IHC immunohistochemistry, HCC hepatocellular carcinoma

Article Snippet: Mouse strain generation and mouse breeding Mice containing Loxp-STOP-Loxp-Fbxl6 (LSL-Fbxl6KI/+, n = 5) were generated by Biocytogen Pharmaceuticals (Beijing, China), and floxed alleles of Fbxl6 (conditional knockout mice of Fbxl6, CKO-Fbxl6KO/+, n = 4) were generated by Cyagen Biosciences Corporation (Guangzhou, China).

Techniques: Immunohistochemistry, Expressing

Journal: Military Medical Research

Article Title: Elevated FBXL6 activates both wild-type KRAS and mutant KRAS G12D and drives HCC tumorigenesis via the ERK/mTOR/PRELID2/ROS axis in mice.

doi: 10.1186/s40779-023-00501-8

Figure Lengend Snippet: Fig. 7 Working model of KRAS/KRASG12D-triggered HCC in response to high FBXL6 expression. FBXL6 elevation facilitates the KRAS/KRASG12D mutation-mediated activation of MEK/ERK/mTOR signaling by promoting K63-linked KRAS/KRASG12D polyubiquitination at the site K128. Hyperactive mTOR increases the expression of PRELID2, leading to HCC tumorigenesis and metastasis in mice. This picture was created by BioRender. FBXL6 F-box and leucine-rich repeat 6, KRAS kirsten rat sarcoma, PRELID2 the proteins of relevant evolutionary and lymphoid interest (PRELI) domain 2, HCC hepatocellular carcinoma, MEK mitogen-activated protein kinase kinase, ERK extracellular signal-regulated kinase, mTOR mammalian target of rapamycin, WT wild-type, Ub ubiquitin, ROS reactive oxygen species

Article Snippet: Mouse strain generation and mouse breeding Mice containing Loxp-STOP-Loxp-Fbxl6 (LSL-Fbxl6KI/+, n = 5) were generated by Biocytogen Pharmaceuticals (Beijing, China), and floxed alleles of Fbxl6 (conditional knockout mice of Fbxl6, CKO-Fbxl6KO/+, n = 4) were generated by Cyagen Biosciences Corporation (Guangzhou, China).

Techniques: Expressing, Mutagenesis, Activation Assay, Ubiquitin Proteomics