| 摘要: |
| ［摘要］ 目的 探究唾液酸结合免疫球蛋白样凝集素15（Siglec-15）通过EGFR/PI3K/AKT信号通路调节肝细胞癌（HCC）细胞的增殖和迁移。方法 利用GEPIA2数据库分析Siglec-15与表皮生长因子受体（EGFR）在HCC组织及癌旁正常组织中的表达，并通过GeneMANIA网站预测二者之间的潜在调控关系。采用实时荧光定量聚合酶链反应（RT-qPCR）和Western blot检测正常肝细胞L02及HCC细胞HepG2、SMMC-7721、BEL-7402中Siglec-15和EGFR的表达。选取HepG2细胞进行后续功能实验，将其分为Siglec-15-NC组、si-Siglec-15组、EGFR-NC组、si-EGFR组、si-Siglec-15+EGFR-NC组、si-Siglec-15+si-EGFR组，并通过RT-qPCR验证各干扰组的转染效率。采用Western blot检测各组细胞中Siglec-15、EGFR/PI3K/AKT信号通路相关蛋白的表达，并利用MTT、Annexin V-FITC/PI、细胞划痕和Transwell实验分别检测转染后Siglec-15、EGFR/PI3K/AKT信号通路单独及联合作用对HCC细胞增殖、凋亡、迁移能力的影响。结果 Siglec-15、EGFR在HCC组织中的基因表达水平显著高于癌旁正常组织（P<0.05），且二者之间存在调控关系。Siglec-15在SMMC-7721、BEL-7402、L02细胞中的表达水平显著低于HepG2细胞（P<0.05），EGFR在SMMC-7721、L02细胞中的表达水平显著低于HepG2细胞（P<0.05）。在HepG2细胞中成功构建了Siglec-15及EGFR的敲低模型。功能实验表明，单独敲低Siglec-15或EGFR均可显著抑制HCC细胞的增殖与迁移能力，并诱导其凋亡。同时敲低Siglec-15与EGFR可通过调控PI3K/AKT信号通路的激活，协同抑制HCC细胞增殖和迁移。结论 Siglec-15与EGFR在HCC中呈高表达，且二者之间存在调控关系。Siglec-15通过EGFR/PI3K/AKT信号通路来调控HCC细胞增殖和迁移。 |
| 关键词: 唾液酸结合免疫球蛋白样凝集素15 EGFR/PI3K/AKT通路 肝细胞癌 增殖 迁移 |
| DOI：10.3969/j.issn.1674-3806.2026.06.10 |
| 分类号:R 735.7 |
| 基金项目:河南省医学科技攻关计划项目（编号：LHGJ20250771） |
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| Siglec-15 regulates the proliferation and migration of hepatocellular carcinoma cells through the EGFR/PI3K/AKT signal pathway |
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HOU Shuting1, GUAN Lulu2, LEI Xiaotian3, TIAN Mengmeng3, GUO Cairu2, WEN Xuejun4, QI Min5
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1.Department of Gastroenterology, Luoyang Central Hospital Affiliated to Zhengzhou University, Zhengzhou 450001, China; 2.Key Laboratory of Individualized Treatment for Liver Cancer, Luoyang Central Hospital Affiliated to Zhengzhou University, Luoyang 471000, China; 3.Third Clinical College of Postgraduate Education, Henan Medical University, Xinxiang 453000, China; 4.Biomanufacturing Laboratory, Virginia Commonwealth University, Charlottesville, Virginia 22903, the United States of America; 5.Henan Provincial Laboratory of Tumor Immunology and Regenerative Medicine, Luoyang 471000, China
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| Abstract: |
| ［Abstract］ Objective To investigate the regulation of the proliferation and migration of hepatocellular carcinoma(HCC) cells by sialic acid-binding immunoglobulin-like lectin 15(Siglec-15) via the EGFR/PI3K/AKT signal pathway. Methods The expressions of Siglec-15 and epidermal growth factor receptor(EGFR) in HCC tissues and adjacent normal tissues were analyzed by using the Gene Expression Profiling Interactive Analysis 2(GEPIA2) database, and the potential regulatory relationship between Siglec-15 and EGFR was predicted by using the GeneMANIA website. The expressions of Siglec-15 and EGFR in normal hepatocyte L02 and HCC cells HepG2, SMMC-7721 and BEL-7402 were detected by using reverse transcription-quantitative real-time polymerase chain reaction(RT-qPCR) and Western blot. The HepG2 cells were used for subsequent functional assays and were assigned to Siglec-15-NC group, si-Siglec-15 group, EGFR-NC group, si-EGFR group, si-Siglec-15+EGFR-NC group and si-Siglec-15+si-EGFR group, and the transfection efficiency in each interference groups was verified by using RT-qPCR. Western blot assay was used to detect the expressions of Siglec-15 and the EGFR/PI3K/AKT signaling pathway-related proteins in the cells of each group. In addition, methyl thiazolyl tetrazolium(MTT), annexin V-fluorescein isothiocyanate(Annexin V-FITC)/propidium iodide(PI) staining, wound-healing assay and Transwell assay were used to evaluate the individual and combined effects of Siglec-15 and the EGFR/PI3K/AKT signaling pathway on HCC cell proliferation, apoptosis, and migration capacities after transfection. Results The gene expression levels of Siglec-15 and EGFR in the HCC tissues were significantly higher than those in the adjacent normal tissues(P<0.05), and there was a regulatory relationship between them. The expression levels of Siglec-15 in the SMMC-7721, BEL-7402 and L02 cells were significantly lower than those in the HepG2 cells(P<0.05). The expression levels of EGFR in the SMMC-7721 and L02 cells were significantly lower than those in HepG2 cells(P<0.05). The stable knockdown models of Siglec-15 and EGFR were successfully established in the HepG2 cells. Functional assays demonstrated that the knockdown of either Siglec-15 or EGFR alone significantly inhibited the proliferation and migration capacities of HCC cells and promoted their apoptosis. The concurrent knockdown of Siglec-15 and EGFR synergistically inhibited the proliferation and migration of HCC cells by regulating activation of the PI3K/AKT signaling pathway. Conclusion Siglec-15 and EGFR are highly expressed in HCC, and there is a regulatory relationship between them. Siglec-15 regulates the proliferation and migration of HCC cells through the EGFR/PI3K/AKT signaling pathway. |
| Key words: Sialic acid-binding immunoglobulin-like lectin 15(Siglec-15) EGFR/PI3K/AKT pathway Hepatocellular carcinoma(HCC) Proliferation Migration |
