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目的 探讨PPFIA4在肝细胞癌组织和HCCLM3细胞中的表达水平及其对肝细胞癌生物学行为的调控。方法 选择生物信息学分析、Western blot法和免疫组化法检测肝细胞癌患者肿瘤组织中PPFIA4表达情况并进行患者预后的相关分析;设计siRNA质粒瞬转敲低HCCLM3细胞中PPFIA4表达,选取划痕愈合实验和Transwell实验检测敲低PPFIA4对HCCLM3细胞迁移、侵袭能力的影响;借助Western blot法检测HCCLM3细胞株转染siRNA质粒后上皮-间充质转化(EMT)相关蛋白标志物的表达变化。结果 PPFIA4在肝细胞癌组织和肝细胞癌细胞(HCCLM3、Li-7、MHCC97H)中高表达;PPFIA4高表达提示患者临床分期较晚,总体生存期(OS)较短;敲低HCCLM3细胞株中PPFIA4表达后,HCCLM3细胞迁移和侵袭能力均下降(P<0.001),并且EMT标志物表达发生变化,上皮细胞标志物E-cadherin的表达增加(P<0.01),而间充质标志物Vimentin、N-cadherin的表达下降(P<0.05、P<0.01)。结论 PPFIA4在肝细胞癌组织和肝癌细胞株中呈高表达并与患者不良预后相关,沉默PPFIA4后能够调控肝癌细胞生物学行为,抑制HCCLM3细胞迁移和侵袭能力,其具体机制可能与EMT相关。
Abstract:Objective To explore the expression level of PPFIA4 in hepatocellular carcinoma tissues and HCCLM3 cells and its regulation of the biological behavior of hepatocellular carcinoma. Methods Bioinformatics analysis, Western blot, and immunohistochemistry were employed to detect the expression of PPFIA4 in tumor tissues of patients with hepatocellular carcinoma and analyze the prognosis of these patients. An siRNA plasmid was designed to knock down the expression of PPFIA4 in HCCLM3 cells. The effects of PPFIA4 knockdown on the migration and invasion abilities of HCCLM3 cells were then evaluated using scratch healing and Transwell assays. Furthermore, Western blot was utilized to detect the expression levels of epithelial-mesenchymal transition(EMT)-related protein markers in the HCCLM3 cell line after transfection with the siRNA plasmid. Results PPFIA4 was highly expressed in hepatocellular carcinoma tissues and hepatocellular carcinoma cells( HCCLM3, Li-7, MHCC97H); the high expression of PPFIA4 indicated that the clinical stage of patients was late and the overall survival(OS) was short. After knocking down the expression of PPFIA4 in HCCLM3 cell line, the migration and invasion ability of HCCLM3 cells decreased(P<0.001) and the expression of EMT markers changed. The expression of epithelial cell marker E-cadherin increased(P<0.01), while the expression of mesenchymal markers Vimentin and N-cadherin decreased(P<0.05, P<0.01). Conclusion PPFIA4 is highly expressed in hepatocellular carcinoma tissues and hepatocellular carcinoma cell lines and is associated with poor prognosis of patients. Silencing PPFIA4 can regulate the biological behavior of hepatocellular carcinoma cells and inhibit the migration and invasion of HCCLM3 cells. The specific mechanism may be related to EMT.
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基本信息:
DOI:10.19405/j.cnki.issn1000-1492.2025.03.005
中图分类号:R735.7
引用信息:
[1]崔浩东,殷济民,郭凯等.PPFIA4基因在肝细胞癌中的表达及作用机制[J].安徽医科大学学报,2025,60(03):414-421.DOI:10.19405/j.cnki.issn1000-1492.2025.03.005.
基金信息:
安徽省重点研究与开发计划人口健康专项项目(编号:202104j07020005); 安徽省高校自然科学研究项目(编号:2023AH010084、2022AH052030)~~