Basic Study
Copyright ©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. May 21, 2024; 30(19): 2553-2563
Published online May 21, 2024. doi: 10.3748/wjg.v30.i19.2553
HepG2.2.15-derived exosomes facilitate the activation and fibrosis of hepatic stellate cells
Yang Gao, Li Li, Sheng-Ning Zhang, Yuan-Yi Mang, Xi-Bing Zhang, Shi-Ming Feng
Yang Gao, Li Li, Sheng-Ning Zhang, Yuan-Yi Mang, Xi-Bing Zhang, Shi-Ming Feng, 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
Author contributions: Gao Y was responsible for the experimental design, execution of experiments, writing, cell culture, and staining; Li L contributed to the experimental design and reviewed the data; Zhang SN participated in writing and reviewing the manuscript; Mang YY provided reagents/materials/analysis tools, conducted nucleic acid extraction, and performed PCR; Zhang XB conducted experiments; Feng SM was involved in conducting experiments, as well as in data collection, analysis, Western blotting (WB), and exosome extraction. All authors critically reviewed and revised the text and approved the final version.
Supported by The Spring City Plan: The High-level Talent Promotion and Training Project of Kunming, No. 2022SCP002; and The Research of Key Techniques and Application of Liver-Kidney Organ Transplantation, No. 202302AA310018.
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board at Affiliated Calmette Hospital of Kunming Medical University & The First Hospital of Kunming.
Institutional animal care and use committee statement: This study does not involve animal research.
Conflict-of-interest statement: All authors declare that they have no conflict of interest.
Data sharing statement: Data is applicable after the approval of the authors.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Li Li, MD, Chief Physician, Department of Hepatobiliary Pancreatic and Vascular Surgery, The Affiliated Calmette Hospital of Kunming Medical University and The First Hospital of Kunming, No. 504 Qingnian Road, Kunming 650011, Yunnan Province, China. kmlili557@163.com
Received: January 13, 2024
Revised: March 5, 2024
Accepted: April 25, 2024
Published online: May 21, 2024
Abstract
BACKGROUND

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.

AIM

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.

METHODS

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.

RESULTS

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.

CONCLUSION

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.

Keywords: Hepatic stellate cells, Liver fibrosis, Exosomes, Small RNA sequencing, HepG2.2.15

Core Tip: This study investigated the effects of exosomes, particularly those derived from HepG2.2.15 cells, on the activation of LX2 stellate cells and the progression of liver fibrosis. Exosomes from HepG2.2.15 cells been found to enhance LX2 cell activation, proliferation, and fibrosis. These exosomes contained 27 differentially expressed microRNAs that target various cellular functions and pathways, including differentiation, signal transduction, and apoptosis regulation. This suggests a significant role for HepG2.2.15-derived exosomes in liver fibrosis, with potential therapeutic implications.