Hydroxylsafflower yellow A modulates tumor vascularization in liver cancer through intercellular communication and...

Fibrosis represents a critical pathological feature in the progression of liver cancer, wherein the activation of hepatic stellate cells (HSCs) plays an important role. Hydroxylsafflower yellow A (HSYA), a chalcone C-glycoside compound, has exhibited hepatoprotective, anti-fibrotic and anti-tumor effects. However, its potential role in preventing fibrosis-associated liver cancer remains unclear.

This study aimed to investigate the effects and underlying mechanisms of HSYA in fibrosis-associated liver cancer through both in vivo and in vitro experimental models.

Single-cell RNA sequencing, bioinformatics analysis, and molecular biology techniques were employed to elucidate the mechanisms and identify the key molecular targets of HSYA in liver cancer. Co-culture experiments involving HSCs-derived exosomes (HSCs-exo) and hepatoma cells were conducted to evaluate the impact of HSYA on intercellular communication within the tumor microenvironment (TME). In the in vivo study, a fibrosis-associated liver cancer mouse model was established using diethylnitrosamine and carbon tetrachloride. Histological staining, immunohistochemistry, western blotting analysis, RT-qPCR, scanning electron microscopy, and fluorescence in situ hybridization were utilized to assess the effects of HSYA on tumor vascularization, including vasculogenic mimicry, angiogenesis, and vascular structural normalization.

miR-29a-3p, a tumor-suppressive miRNA, was significantly downregulated in liver cancer patients and exerted anti-tumor effects predominantly by modulating of extracellular matrix (ECM)-receptor interactions and the PI3K/AKT signaling pathway. HSYA effectively upregulated miR-29a-3p expression in both HSCs-exo and tumor tissues, thereby attenuating vasculogenic mimicry and abnormal angiogenesis in hepatoma cells as well as in the fibrosis-associated liver cancer mouse model through inhibition of the Col-IV/ITGB1/FAK and PI3K/AKT signaling pathways. Co-localization analysis of miR-29a-3p, VEGFA, and vimentin further demonstrated that HSYA increased miR-29a-3p expression while concurrently suppressing HSCs activation, trans-differentiation, and tumor vascularization. Furthermore, the study further demonstrated that HSYA partially regulated ECM-receptor interactions and the PI3K/AKT signaling pathway through regulation of HSCs-exo following transfection with a miR-29a-3p inhibitor.

Understanding the dynamic variations in the TME during the progression from liver fibrosis to liver cancer, our findings suggested that the intercellular communication and ECM-receptor interactions regulated by HSYA may represent a promising therapeutic strategy for the treatment of fibrosis-associated liver cancer.