Hepatocellular carcinoma remains a leading cause of cancer-related mortality worldwide. Although lenvatinib is widely deployed as a frontline systemic therapy for advanced disease, its clinical utility is frequently subverted by primary or adaptive non-responsiveness. Evasion of ferroptosis, an iron-dependent form of regulated cell death driven by lipid peroxidation, has emerged as a key survival mechanism for tumor cells under therapeutic stress. While the RNA-binding protein RBMS1 is known to participate in oncogenic progression and ferroptotic regulation across several malignancies, whether it modulates lenvatinib sensitivity in hepatocellular carcinoma by intersecting with the SLC7A11-dependent antioxidant pathway has not been established. Expression profiles of RBMS1 and SLC7A11 were interrogated in paired clinical tissue samples and public transcriptomic datasets. Stable lentiviral overexpression or short hairpin RNA-mediated knockdown was executed in HepG2 and Huh7 cells to modulate candidate gene abundance. Cellular phenotypes, comprising viability, clonogenicity, migration, invasion, and drug sensitivity, were mapped via cell counting kit-eight, colony formation, wound healing, and transwell migration assays. Ferroptotic kinetics and oxidative shifts were tracked by measuring intracellular reactive oxygen species and lipid peroxidation using fluorescent probes. Finally, a subcutaneous xenograft model was established to validate the synergistic potential of RBMS1 depletion with lenvatinib therapy in vivo. In clinical specimens, SLC7A11 was markedly upregulated and tied to shortened overall survival, while RBMS1 expression exhibited a strong upward trend that correlated robustly with SLC7A11 abundance in broader transcriptomic cohorts. Functionally, overexpressing RBMS1 accelerated malignant behaviors, expanded SLC7A11 protein levels, neutralized reactive oxygen species, and shielded cells from lipid peroxidation, ultimately lowering lenvatinib sensitivity. Conversely, silencing RBMS1 yielded opposite phenotypes, aggravating ferroptotic oxidative damage and amplifying lenvatinib responsiveness. Epistatic rescue experiments demonstrated that SLC7A11 knockdown completely abrogated the anti-ferroptotic defense and drug tolerance conferred by RBMS1 hyperactivation. In vivo animal models corroborated these dynamics, showing that RBMS1 knockdown significantly potentiated the antitumor efficacy of lenvatinib. These findings position RBMS1 as a novel upstream post-transcriptional regulator of SLC7A11-dependent ferroptosis susceptibility and lenvatinib responsiveness in hepatocellular carcinoma. By reinforcing the SLC7A11 antioxidant network, RBMS1 shields malignant cells from drug-induced oxidative catastrophe. Targeting this newly uncovered post-transcriptional axis represents a viable therapeutic strategy to overcome lenvatinib tolerance in advanced disease.

BACKGROUND:Hepatocellular carcinoma (HCC) is one of the most common malignant tumors, characterized by high recurrence and metastasis rates. Studies have demonstrated that exosomes, as signaling mediators between tumor cells and their microenvironment, play crucial roles in tumor cell migration and invasion. S100A4, a known metastasis-associated protein, has been identified as a key factor in promoting cancer cell migration and epithelial-mesenchymal transition (EMT). METHODS:To investigate the relationship between S100A4 and high metastasis in HCC, we performed analyses using TCGA-HCC data and clinical samples. Stable HCC cell lines overexpressing S100A4 were established to evaluate S100A4 expression levels among various HCC cell types. The migration and invasion capabilities of these cells were assessed using wound healing and Transwell assays. Additionally, Western blotting, qPCR, and immunohistochemistry (IHC) were employed to detect changes in EMT-related markers and NMIIA expression following S100A4 overexpression. RESULTS:Data from TCGA and IHC analyses revealed that S100A4 is highly expressed in HCC patients and that its elevated expression correlates with lymph node metastasis and advanced tumor stage. High S100A4-expressing (S100A4^H) HCC cells (JHH-1 and Li-7) exhibited enhanced migratory and invasive capabilities, whereas low S100A4-expressing cells (Huh-7 and Hep G2) displayed weaker metastatic characteristics. Exosomes derived from S100A4^H cells also showed elevated levels of S100A4. Overexpression of S100A4 in HCC cells promoted cell migration, invasion, and viability. Moreover, exosomes derived from S100A4^H cells significantly enhanced the migration and invasion of co-cultured HCC cells. Mechanistically, S100A4 overexpression significantly downregulated E-cadherin expression while upregulating Twist1, N-cadherin, and Vimentin levels. Notably, S100A4 formed a complex with NMIIA in HCC cells, and NMIIA silencing suppressed S100A4-induced changes in EMT-related protein expression. CONCLUSION:S100A4 promotes HCC cell migration, invasion, and metastasis by activating the EMT process via NMIIA, potentially through exosome-mediated signaling. S100A4 may serve as a potential biomarker for predicting EMT occurrence, disease progression, and prognosis in HCC patients.

Purpose: Due to resistance to gemcitabine (GEM), patients with pancreatic cancer (PC) usually have poor prognosis and low survival rate. The purpose of our research was to explore the impact of exosome PPP3CB on GEM resistance in PC, and concurrently analyze the regulatory role of the miR-298/STAT3 signaling pathway. Methods: Exosomes isolated from PC cells were verified by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and western blotting (WB). The interaction between PPP3CB and miR-298 was verified using dual-luciferase reporter gene assay, followed by evaluation of cell growth and death using CCK8 assay, EdU staining, and flow cytometry. Results: Increased PPP3CB expression was observed in GEM-resistant PC cells. Exosomes from PC cells and GEM-resistant PC cells were successfully extracted by ultra-high speed centrifugation. Confocal microscopy showed internalization of fluorescein amide (FAM)-labeled GEM-resistant exosomes by PC cells. PPP3CB enhanced the proliferation of GEM-resistant PC cells and inhibited their apoptosis, whereas down-regulation of PPP3CB promoted the death of PC cells and inhibited the proliferation of GEM-resistant PC cells, and enhance the susceptibility of PC cells to GEM. Additionally, PPP3CB positively regulated STAT3 expression in PC cells by down-regulating miR-298, thus promoting the growth and inhibiting the death of PC cells. Conclusion: PC cell-derived exosome PPP3CB enhances STAT3 expression by downregulating miR-298, stimulating cell growth, and suppressing cell death, thereby increasing the resistance of PC cells to GEM.

Hepatocellular carcinoma (HCC) is a common liver cancer that accounts for 90% of cases. Doxorubicin exhibits a broad spectrum of antitumor activity and is one of the most active agents in HCC. WW domain-containing protein 2 (WWP2) is highly expressed in HCC tissues and activates protein kinase B (AKT) signaling pathway to enhance tumor metastasis. However, the role of WWP2 in the glycolysis and antitumor effects of doxorubicin and the epigenetic alterations of WWP2 in HCC remain to be elucidated. The levels of WWP2 and N6-methyladenosine methyltransferase-like 3 (METTL3) in clinical samples and cells were investigated. WWP2 were silenced or overexpressed to study the role of WWP2 in regulating cell proliferation, colony formation, and glycolysis. RNA immunoprecipitation was performed to test m6 A levels. Quantitative reverse-transcription polymerase chain reaction (RT-PCR) and Western blot were used to measure mRNA and protein, respectively. WWP2 silencing inhibits cell proliferation, colony formation, and glycolysis, while WWP2 overexpression has the inverse effects via the AKT signaling pathway. Silencing WWP2 enhances doxorubicin's antitumor effect, while WWP2 overexpression suppresses doxorubicin's antitumor effect. Data also support that METTL3 mediates WWP2 m6A modification, and m6A reader, IGF2BP2, binds to the methylated WWP2 to promote the stability of WWP2, leading to upregulation of WWP2. METTL3 mediates WWP2 m6A modification, which can be recognized and bound by IGF2BP2 to increase the stability of WWP2, leading to WWP2 overexpression which inhibits the antitumor effects of doxorubicin through METTL3/WWP2/AKT/glycolysis axis.

目的 探讨WWP的E3泛素连接酶2(ww domain-containing protein 2,WWP2)沉默对肝癌细胞系黏附、侵袭和迁移的影响作用.方法 通过RNA沉默技术降低肝癌细胞系的WWP2水平,用Transwell法检测细胞迁移和侵袭,CCK-8法检测细胞的增殖情况,Western blot法检测caspase7、caspase8、Bcl-2、Bax、PTEN、p-Akt及Akt的蛋白水平.结果 沉默WWP2后,肝癌细胞BEL-7404和Huh7的WWP2mRNA和蛋白的表达水平明显下降(P<0.001),BEL-7404细胞和Huh7细胞的凋亡相关标志物caspase7、caspase8及Bax的蛋白水平表达都显著升高(P<0.01)、PTEN的蛋白水平升高(P<0.01),Bcl-2、p-Akt的蛋白水平显著下降(P<0.01),肝癌细胞BEL-7404和Huh7的细胞活力、黏附能力、侵袭能力和迁移能力都明显减弱(P<0.001).结论 沉默WWP2可抑制肝癌BEL-7404细胞和Huh7细胞增殖、黏附、侵袭和迁移,并促进细胞凋亡,其作用机制可能与PTEN/Akt信号通路相关.

One of the most prominent characteristics of hepatic ischemia-reperfusion injury (HI/R) is an intense inflammatory reaction, which plays a key role in inflammatory injury induced by ischemia-reperfusion. Nucleotide-binding oligomerization domain-containing protein (NOD-), leucine-rich repeat (LRR), and pyrin domains-containing protein 3 (NLRP3) are involved in the inflammatory injury of ischemia-reperfusion as an important pattern recognition receptor for innate immunity. G protein-coupled receptor 30 (GPR30) is a newly identified as 7-transmembrane G protein-coupled receptor and can be activated by many stimulations including estrogen. The current study aims to explore whether GPR30 agonist (G1) can alleviate hepatic ischemia-reperfusion injury HI/R by inhibiting NLRP3. An induced HI/R rat model was generated, blood and liver samples were gathered and subjected to histological examination, biochemical assays, Western blot assays, and qRT-PCR. Our results indicated GPR30 agonist (G1) pretreatment or NLRP3 silencing significantly decreased the serum levels of Interleukin 1 beta (IL-1 beta), alanine aminotransferase (ALT) and aspartate aminotransferase, improved histological alterations and hepatocyte apoptosis. Moreover, G1 pretreatment or NLRP3 silencing downregulated the protein level of Caspase-1 and pro-Interleukin 1 beta (pro-IL-1 beta) while G1 pretreatment upregulated the expression of GPR30 (p < 0.05). In conclusion, the salutary effects of GPR30 agonists on HI/R are mediated at least in part through downregulating NLRP3 expression. GPR30 may be used as a therapy target of HI/R.

Primary liver cancer is one of the most common and aggressive human malignancies worldwide. As numerous studies have revealed that WW domain containing E3 Ub‑protein ligase 2 (WWP2) exerts cancer‑specific functions, the present study assessed the role of WWP2 in liver cancer. WWP2 was revealed to be significantly overexpressed in liver cancer tissues compared with paired normal tissues at the mRNA as well as at the protein level. Furthermore, small interfering RNA-mediated WWP2 knockdown in liver cancer cell lines was demonstrated to inhibit cell proliferation, cause cell cycle arrested in G1 phase and to induce apoptosis as revealed by a Cell Counting Kit-8 assay and flow cytometric analysis. In addition, western blot analysis revealed that WWP2 knockdown significantly increased the expression of apoptosis-associated markers caspase‑7, caspase‑8 and B-cell lymphoma 2 (Bcl-2)-associated X in liver cancer cell lines, while Bcl‑2 was significantly decreased. In conclusion, the present study suggested that WWP2 may exert important functions in the over‑proliferation and evasion of apoptosis of liver cancer, likely through regulating the expression of apoptosis-associated markers. Furthermore, WWP2 may represent a novel diagnostic marker and molecular therapeutic target for liver cancer.

The aim of the present study is to investigate the effect and mechanism of recombinant human acidic fibroblast growth factor (rhaFGF) on skin wound healing of diabetic rats. SD rats were randomly divided into three groups: control, diabetes and rhaFGF groups. Skin wound tissues of rats in the three groups were respectively arranged on days 3 and 7 after establishment of skin wound of diabetic rats. Immunohistochemistry analyses were performed to evaluate the effect of rhaFGF on expression levels of PCNA and CD34 in diabetic rats. Real-time PCR and Western blot assays were used to examine the expression levels of Bcl-2, Bax, TGF-beta, CXCL-1, CXCL-2, p-ERK1/2 and p-STAT3 in wound tissues. The results showed that rhaFGF could significantly increased expression levels of PCNA, CD34, TGF-beta, CXCL-1, CXCL-2, p-ERK1/2, p-STAT3 and the ratio of Bcl-2/Bax in wound tissues. The preliminary mechanism that rhaFGF accelerates wound healing of diabetic rats is possibly associated with the promotion of fibroblasts proliferation and angiogenesis through ERK1/2 and STAT3 pathways.

OBJECTIVE:To discuss the efficacy of contrast-enhanced ultrasound (CEUS) for diagnosis of small hepatocellular carcinoma (HCC) by pooling the open published data. METHODS:A comprehensive publication electronic search was performed by reviewers in the databases of PubMed, Embase, Web of Science, and China National Knowledge Infrastructure. The open published studies about CEUS for small HCC diagnosis were collected. The sensitivity, specificity, positive likelihood ratio (+LR), negative likelihood ratio (-LR), and diagnostic odds ratio (DOR) were pooled by stata 12.0 software. RESULTS:A total of 16 studies were included in the present study. The sensitivity, specificity, +LR, and -LR were aggregated by random effects model because of significant heterogeneity (I2 >50.0%). However, DOR was pooled by fixed effects model without significant heterogeneity (I2 <50.0%). The aggregate sensitivity was 0.86 (95% confidence interval [CI]: 0.79-0.91); the aggregate specificity was 0.87 (95% CI: 0.75-0.94); the aggregate +LR and -LR were 7.06 (95% CI: 1.64-30.36) and 0.20 (95% CI: 1.64-30.36), respectively. The DOR was 33.71 (95% CI: 20.34-55.88); the area under the receiver operating characteristic was 0.93 (95% CI: 0.90-0.95). There was significant publication bias tested by funnel plot and line regression test (t = 2.29, P < 0.05). CONCLUSION:With the present evidence, CEUS is useful for diagnosis of small HCC with relatively high sensitivity and specificity.

The role and clinical implication of the WWP2 E3 ubiquitin ligase in liver cancer are poorly understood. In the current study, we investigated the expression level of WWP2 and its functions in cell adhesion, invasion, and migration in liver cancer. We used real-time PCR to detect the expression of WWP2 in liver cancer and adjacent samples from the People’s Hospital of Lishui and also analyzed The Cancer Genome Atlas (TCGA) RNA-seq data by bioinformatics. Migration and invasion were detected by transwell analysis. We detected a strong WWP2 expression in tumor tissues of the People’s Hospital of Lishui, and the survival rate was significantly higher in patients with lower WWP2-expressing tumors. WWP2 small hairpin RNA (shRNA) lentivirus stably infected cells (shWWP2), Huh7, showed slower growth speed compared with scramble control-infected cells in a xenograft mouse model. Knockdown of WWP2 Huh7 and BEL-7404 cells demonstrated a reduction in adhesion, invasion, and migration. Gene set enrichment analysis (GSEA) showed that WWP2 is positively correlated to cancer-related pathways including the chemokine signaling pathway. WWP2 also regulated MMP-9, caspase-9, CXCR3, and CCR5 expression in liver cancer cells. In addition, knockdown of CXCR3 and CCR5 significantly inhibited cell proliferation, adhesion, invasion, and migration in Huh7 and BEL-7404 cells. Our data suggest that targeting of WWP2 may be a therapeutic strategy for liver cancer treatment.