scRNA-Seq Analysis Revealed CAFs Regulating HCC Cells via PTN Signaling

J Hepatocell Carcinoma. 2024 Nov 18:11:2269-2281. doi: 10.2147/JHC.S493675. eCollection 2024.

Abstract

Background: Cancer-associated fibroblasts (CAFs) play a pivotal role in shaping the microenvironment of hepatocellular carcinoma (HCC). However, the mechanisms through which CAFs influence the progression of HCC remain incompletely understood.

Methods: Single-cell RNA sequencing datasets (GSE158723 and GSE112271) were retrieved from the Gene Expression Omnibus (GEO) database at the National Center for Biotechnology Information (NCBI) and analyzed using R software. Our analysis suggested that CAFs may promote liver cancer cell development, possibly through the interaction of pleiotrophin (PTN) and syndecan-2 (SDC2). Clinical samples from HCC patients were collected and processed into frozen sections and single-cell suspensions for Masson staining, immunofluorescence staining, and flow cytometry. Additionally, Huh7 liver cancer cells and LO2 normal liver cells were cultured and subjected to immunofluorescence assays using cell slides.

Results: The proportion of CAFs in cancerous tissues was higher than in adjacent non-cancerous tissues, and pleiotrophin (PTN) expression was elevated in cancer tissues compared to adjacent tissues. These findings aligned with the results of the single-cell RNA sequencing (scRNA-seq) analysis. Furthermore, SDC2 expression was significantly upregulated in Huh7 liver cancer cells compared to LO2 normal liver cells.

Discussion: This study suggests that CAFs may contribute to HCC progression via the PTN/SDC2 signaling pathway. Our findings provide deeper insights into the interactions between CAFs and HCC cells within the tumor microenvironment (TME).

Keywords: CAFs; HCC; PTN; TME; scRNA-seq.

Grants and funding

This work was supported by the National Natural Science Foundation of China (#82160589), Guangxi Science and Technology Major Project (#GuikeAA23073013), The Natural Science Foundation of Guangxi Zhuang Autonomous Region (#2024GXNSFAA010126), Nanning Qingxiu District Science and Technology Bureau Key Research and Development Plan (#2020035), Guangxi Key Laboratory of Early Prevention and Treatment for Regional High-Frequency Tumors (#GXK201604), Key Laboratory of Basic Research on Regional Diseases (Guangxi Medical University), Education Department of Guangxi Zhuang Autonomous Region (#GXQYJB2023001), and Guangxi Medical and Health Program for Promotion (#s2022140).