Background: Hepatocellular carcinoma (HCC) remains a global challenge due to its high morbidity and mortality rates as well as poor response to treatment. The communication between tumor-derived elements and stroma plays a critical role in facilitating cancer progression of HCC. Exosomes are small extracellular vesicles (EVs) that are released from the cells upon fusion of multivesicular bodies with the plasma membrane. There is emerging evidence indicating that exosomes play a central role in cell-to-cell communication. Much attention has been paid to exosomes since they are found to transport bioactive proteins, messenger RNA (mRNAs) and microRNA (miRNAs) that can be transferred in active form to adjacent cells or to distant organs. However, the mechanisms underlying such cancer progression remain largely unexplored.Methods: Exosomes were isolated by differential ultracentrifugation from conditioned medium of HCC cells and identified by electron microscopy and Western blotting analysis. Hepatic stellate cells (HSCs) were treated with different concentrations of exosomes, and the activation of HSCs was analyzed by Western blotting analysis, wound healing, migration assay, Edu assay, CCK-8 assay and flow cytometry. Moreover, the different miRNA levels of exosomes were tested by real-time quantitative PCR (RT-PCR). The angiogenic ability of activated HSCs was analyzed by qRT-PCR, CCK-8 assay and tube formation assay. In addition, the abnormal lipid metabolism of activated HSCs was analyzed by Western blotting analysis and Oil Red staining. Finally, the relationship between serum exosomal miRNA-21 and prognosis of HCC patients was evaluated.Results: We showed that HCC cells exhibited a great capacity to convert normal HSCs to cancer-associated fibroblasts (CAFs). Moreover, our data revealed that HCC cells secreted exosomal miRNA-21 that directly targeted PTEN, leading to activation of PDK1/AKT signaling in HSCs. Activated CAFs further promoted cancer progression by secreting angiogenic cytokines, including VEGF, MMP2, MMP9, bFGF and TGF-β. Clinical data indicated that high level of serum exosomal miRNA-21 was correlated with greater activation of CAFs and higher vessel density in HCC patients.Conclusions: Intercellular crosstalk between tumor cells and HSCs was mediated by tumor-derived exosomes that controlled progression of HCC. Our findings provided potential targets for prevention and treatment of live cancer.

We present NY-07, an antifolate dye conjugate capable of intrasurgical discrimination between inflammatory and cancerous tissues. This capability addresses a longstanding problem in fluorescence-guided surgery, where the current FDA-approved probes exhibit false-positive rates of 25–68% due to nonspecific accumulation in inflammatory tissue, leading to unnecessary tissue resection and increased surgical morbidity. NY-07 selectively targets folate receptor α (FRα) overexpressed in tumors while exhibiting 8-fold reduced affinity for folate receptor β (FRβ) predominantly found on inflammatory macrophages (K _d = 61.67 nM vs 486.9 nM). In mouse models, NY-07 achieved a tumor-to-background ratio of 3.23 ± 0.28 with fluorescence signals in tumor tissue significantly exceeding inflammatory areas (p < 0.01). The probe detected submillimeter cancer lesions while avoiding false signals in pneumonia and arthritis models. Co-localization studies using immunohistochemical staining and fluorescence microscopy confirmed that FRα-positive areas in tumor tissue exhibited strong fluorescence intensity, while FRβ-positive areas showed a minimal signal. NY-07 maintained diagnostic imaging windows exceeding 12 days and demonstrated acceptable safety profiles in Phase I clinical trials (CTA: CXHL2401187), having received IND approval from both FDA and NMPA. These results position NY-07 as a strong clinical candidate with the potential to improve surgical precision and reduce false-positive resections.