Department of Hepatobiliary Pancreatic and Vascular Surgery, The Affiliated Calmette Hospital of Kunming Medical University and The First Hospital of Kunming, Kunming 650011, Yunnan Province, China.
World J Gastroenterol. 2024 May 21;30(19):2553-2563. doi: 10.3748/wjg.v30.i19.2553.
The role of exosomes derived from HepG2.2.15 cells, which express hepatitis B virus (HBV)-related proteins, in triggering the activation of LX2 liver stellate cells and promoting liver fibrosis and cell proliferation remains elusive. The focus was on comprehending the relationship and influence of differentially expressed microRNAs (DE-miRNAs) within these exosomes.
To elucidate the effect of exosomes derived from HepG2.2.15 cells on the activation of hepatic stellate cell (HSC) LX2 and the progression of liver fibrosis.
Exosomes from HepG2.2.15 cells, which express HBV-related proteins, were isolated from parental HepG2 and WRL68 cells. Western blotting was used to confirm the presence of the exosomal marker protein CD9. The activation of HSCs was assessed using oil red staining, whereas DiI staining facilitated the observation of exosomal uptake by LX2 cells. Additionally, we evaluated LX2 cell proliferation and fibrosis marker expression using 5-ethynyl-2'-deoxyuracil staining and western blotting, respectively. DE-miRNAs were analyzed using DESeq2. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were used to annotate the target genes of DE-miRNAs.
Exosomes from HepG2.2.15 cells were found to induced activation and enhanced proliferation and fibrosis in LX2 cells. A total of 27 miRNAs were differentially expressed in exosomes from HepG2.2.15 cells. GO analysis indicated that these DE-miRNA target genes were associated with cell differentiation, intracellular signal transduction, negative regulation of apoptosis, extracellular exosomes, and RNA binding. KEGG pathway analysis highlighted ubiquitin-mediated proteolysis, the MAPK signaling pathway, viral carcinogenesis, and the toll-like receptor signaling pathway, among others, as enriched in these targets.
These findings suggest that exosomes from HepG2.2.15 cells play a substantial role in the activation, proliferation, and fibrosis of LX2 cells and that DE-miRNAs within these exosomes contribute to the underlying mechanisms.
表达乙型肝炎病毒（HBV）相关蛋白的 HepG2.2.15 细胞衍生的外泌体在触发 LX2 肝星状细胞（HSC）激活并促进肝纤维化和细胞增殖中的作用尚不清楚。重点是理解这些外泌体中差异表达的 microRNAs（DE-miRNAs）之间的关系和影响。
阐明来自表达 HBV 相关蛋白的 HepG2.2.15 细胞的外泌体对 HSC LX2 激活和肝纤维化进展的影响。
从亲本 HepG2 和 WRL68 细胞中分离出表达 HBV 相关蛋白的 HepG2.2.15 细胞衍生的外泌体。Western blot 用于确认外泌体标记蛋白 CD9 的存在。油红染色用于评估 HSCs 的激活，而 DiI 染色有助于观察 LX2 细胞摄取外泌体。此外，我们使用 5-乙炔基-2'-脱氧尿苷染色和 Western blot 分别评估 LX2 细胞增殖和纤维化标记物的表达。使用 DESeq2 分析 DE-miRNA。基因本体论（GO）和京都基因与基因组百科全书（KEGG）途径用于注释 DE-miRNA 的靶基因。
发现 HepG2.2.15 细胞的外泌体诱导 LX2 细胞激活并增强其增殖和纤维化。HepG2.2.15 细胞外泌体中共有 27 个 miRNA 表达差异。GO 分析表明，这些 DE-miRNA 靶基因与细胞分化、细胞内信号转导、细胞凋亡的负调控、细胞外外泌体和 RNA 结合有关。KEGG 途径分析突出了泛素介导的蛋白水解、MAPK 信号通路、病毒致癌作用和 Toll 样受体信号通路等在这些靶基因中的富集。
这些发现表明 HepG2.2.15 细胞的外泌体在 LX2 细胞的激活、增殖和纤维化中起重要作用，并且这些外泌体中的 DE-miRNA 有助于潜在的机制。
