目的:Perilipin1(PLIN1)是一种必需的脂滴表面蛋白,通过调节能量平衡和脂质代谢来参与细胞生命活动。PLIN1已被证明与许多肿瘤类型的发展密切相关。这项工作的目的是阐明PLIN1在肝细胞癌(HCC)中的临床病理意义,以及它对肝癌细胞生物学功能的影响,并调查其中的潜在机制。
方法:收集公共高通量RNA微阵列和RNA测序数据,以检查肝癌患者的PLIN1水平和临床意义。进行免疫组织化学(IHC)和实时定量逆转录聚合酶链反应(RT-qPCR)以评估PLIN1在HCC中的表达水平和临床病理相关性。然后,用过表达PLIN1的慢病毒转染SK和Huh7细胞。CCK8测定,伤口愈合试验,transwell分析,和流式细胞术分析,以探讨PLIN1过表达对肝癌细胞增殖的影响,迁移,入侵,和细胞周期分布。最终,进行基因本体论(GO)功能注释和京都基因和基因组百科全书(KEGG)途径分析,以基于HCC差异表达基因和PLIN1共表达基因研究PLIN1在HCC进展中的潜在机制。
结果:PLIN1在HCC组织中明显下调,这与HCC患者的预后明显较差有关。此外,PLIN1过表达抑制细胞增殖,迁移,SK和Huh7细胞的体外侵袭,以及将HCC细胞周期阻滞在G0/G1期。更重要的是,与能量转化相关的生物过程,脂质代谢,和细胞周期信号通路是三种最富集的分子机制。
结论:本研究表明,PLIN1下调与HCC患者的不良预后相关,并通过促进细胞增殖加速HCC进展,迁移,和转移,以及肝癌中脂质代谢相关通路的调节机制。
OBJECTIVE: Perilipin 1 (PLIN1) is an essential lipid droplet surface protein that participates in cell life activities by regulating energy balance and lipid metabolism. PLIN1 has been shown to be closely related to the development of numerous tumor types. The purpose of this work was to elucidate the clinicopathologic significance of PLIN1 in hepatocellular carcinoma (HCC), as well as its impact on the biological functions of HCC cells, and to investigate the underlying mechanisms involved.
METHODS: Public high-throughput RNA microarray and RNA sequencing data were collected to examine PLIN1 levels and clinical significance in patients with HCC. Immunohistochemistry (IHC) and real-time quantitative reverse transcription polymerase chain reaction (RT‒qPCR) were conducted to assess the expression levels and the clinicopathological relevance of PLIN1 in HCC. Then, SK and Huh7 cells were transfected with a lentivirus overexpressing PLIN1. CCK8 assay, wound healing assay, transwell assay, and flow cytometric analysis were conducted to explore the effects of PLIN1 overexpression on HCC cell proliferation, migration, invasion, and cell cycle distribution. Ultimately, Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to investigate the underlying mechanisms of PLIN1 in HCC progression based on HCC differentially expressed genes and PLIN1 co-expressed genes.
RESULTS: PLIN1 was markedly downregulated in HCC tissues, which correlated with a noticeably worse prognosis for HCC patients. Additionally, PLIN1 overexpression inhibited the proliferation, migration, and invasion of SK and Huh7 cells in vitro, as well as arresting the HCC cell cycle at the G0/G1 phase. More significantly, energy conversion-related biological processes, lipid metabolism, and cell cycle signalling pathways were the three most enriched molecular mechanisms.
CONCLUSIONS: The present study revealed that PLIN1 downregulation is associated with poor prognosis in HCC patients and accelerated HCC progression by promoting cellular proliferation, migration, and metastasis, as well as the mechanisms underlying the regulation of lipid metabolism-related pathways in HCC.