肝星状细胞通过全反式维甲酸介导肝癌细胞凋亡的作用及分子机制研究OA

Objective:To investigate whether hepatic stellate cells(HSCs)induce apoptosis in hepatocellular carcinoma(HCC)cells by secreting all-trans retinoic acid(ATRA),and to elucidate the underlying molecular mecha-nisms.Methods:An in vitro co-culture system was estab-lished using LX2 cells and three HCC cell lines(HepG2,PLC/PRF/5,and MHCC97H).Apoptosis of HCC cells was detected by flow cytometry(FCM).Changes in ATRA concen-trations in the supernatant of LX2 cell cultures were measured with a microplate reader.Meanwhile,CCK-8 assay and FCM were used to detect cell viability and apoptosis of HepG2 cells after being treated with ATRA.RNA sequencing(RNA-seq)was performed on HepG2 cells subjected to LX2 co-culture or exogenous ATRA intervention.Bioinformatics analysis was then conducted to identify differentially expressed genes(DEGs),core signaling pathways,and pivotal genes under both condi-tions.The screened candidate key genes were further validated by RT-qPCR.Results:FCM results showed that co-culture with LX2 cells significantly promoted apoptosis in HepG2,PLC/PRF/5,and MHCC97H cells.LX2 cell supernatant con-tained ATRA,whose concentration increased gradually in a time-dependent manner.CCK-8 and FCM results indicated that ATRA treatment markedly inhibited cell viability and induced apoptosis in HepG2 cells.RNA-seq analysis identified 304 co-regulated DEGs in HepG2 cells upon LX2 co-culture and exogenous ATRA treatment.Functional enrichment analysis demon-strated that these genes were mainly enriched in pivotal signaling pathways,including apoptosis,TNF signaling pathway,and HIF-1 signaling pathway.A total of 12 apoptosis-related co-regulated genes were further screened out.RT-qPCR results con-firmed significant upregulation of MEIS3,LGALS3,INHBB,and ICAM1 expression.Conclusion:This study demonstrates that HSCs promote HCC cell apoptosis via secreting ATRA;the underlying mechanism involves transcriptomic reprogramming and upregulation of key apoptotic genes.