目的：探讨CXCL5/CXCR2/VEGF通路在HBV相关肝癌血管生成过程中的作用及其机制。方法：收集2022年10月 至2022年12月间在广西中医药大学附属瑞康医院入院的 10 例 HBV-DNA阳性患者的外周血标本[其中5例肝细胞癌（HCC） 和5例肝硬化（LC）]，用高通量RNA测序技术检测并筛选差异表达mRNA，用GO富集和KEGG通路分析这些差异表达基因的生 物学功能和相关信号通路。常规培养HCC细胞HepG2，用转染试剂将C-X-C基序趋化因子配体5（CXCL5）过表达质粒和对照质 粒转染至HepG2细胞中，分为对照组，空载组，250 ng/mL和520 ng/mL CXCL5过表达质粒组。用CCK-8法、ELISA、血管生成实 验、qPCR 法、WB 法和免疫荧光染色技术分别检测过表达 CXCL5 对 HepG2 细胞增殖能力、VEGF 分泌、成管能力、CXCL5、 CXCR2、VEGF mRNA及其蛋白表达，以及VEGF表达的影响。结果：RNA测序结果显示，HCC和LC患者外周血中 mRNA表达 存在显著差异。GO富集分析发现，这些差异mRNA的相关基因主要参与细胞发育过程的调控、G蛋白偶联受体活性、细胞外区 域的组成和信号受体结合。KEGG通路分析发现，这些异表达基因可能参与细胞因子与受体相互作用通路、cAMP信号通路等与 癌症作用机制相关的通路、神经活性配体与受体相关作用的通路、钙相关信号通路。过表达CXCL5可明显促进HepG2细胞的增 殖能力（P < 0.05）、VEGF的分泌（P < 0.05）、人脐静脉内皮细胞（HUVEC）的成管能力（P < 0.05）、CXCL5、CXCR2和VEGF mRNA 和蛋白的表达（P < 0.05或P < 0.01），以及增强HepG2细胞中VEGF的表达。结论：CXCL5可能通过CXCL5/CXCR2/VEGF轴促 进HepG2细胞的增殖和HUVEC的成管能力。

Objective: To explore the role and mechanism of CXC chemokine ligand 5/CXC receptor 2/ vascular endothelial growth factor (CXCL5/CXCR2/VEGF) pathway in the angiogenesis of hepatitis B virus (HBV) -related hepatocellular carcinoma (HCC). Methods: Peripheral blood samples were collected from 10 HBV-DNA positive patients admitted to Ruikang Hospital Affiliated to Guangxi University of Traditional Chinese Medicine between October 2022 and December 2022, including 5 cases of HCC and 5 cases of liver cirrhosis (LC). High-throughput RNA sequencing was used to detect and screen differentially expressed mRNAs. GO enrichment and KEGG pathway analysis were conducted to explore the biological functions and related signaling pathways of these differentially expressed genes. HCC HepG2 cells were conventionally cultured, and CXCL5 overexpression plasmid and control plasmid were transfected into HepG2 cells using transfection reagents. The cells were divided into a control group, an empty load group, and two CXCL5 overexpression groups (250 ng/mL and 520 ng/mL). The effects of CXCL5 overexpression on HepG2 cell proliferation (CCK-8 method), VEGF secretion (ELISA), tube formation ability (angiogenesis assay), mRNA and protein expression of CXCL5, CXCR2, VEGF (qPCR, Western blot), as well as VEGF expression immunofluorescence intensity were analyzed. Results: RNA sequencing results showed significant differences in mRNA expression in peripheral blood between HCC and LC patients. GO enrichment analysis revealed that these differentially expressed genes were mainly involved in cell development regulation, G proteincoupled receptor activity, extracellular region components, and receptor-ligand binding. KEGG pathway analysis revealed that these differentially expressed genes might participate in pathways related to the cytokine-receptor interactions, cAMP signaling and other cancer related pathways, neuroactive ligand-receptor interactions, and calcium signaling pathways. Overexpression of CXCL5 significantly promoted HepG2 cell proliferation (P < 0.05), VEGF secretion (P < 0.05), tube formation in human umbilical vein endothelial cells (HUVECs) (P < 0.05), and mRNA and protein expression of CXCL5, CXCR2, and VEGF (P < 0.05 or P < 0.01), as well as enhanced the immunofluorescence intensity of VEGF in HepG2 cells. Conclusion: CXCL5 may promote the proliferation of HepG2 cells and HUVEC angiogenesis through the CXCL5/CXCR2/VEGF axis.

广西岐黄学者建设项目；广西中医药大学岐黄工程高层次人才培育项目（No.2021007）；广西中医药大学2023年研究生教育创新计划 项目（No.YCBXJ2023032）