广西医学

目的探讨磷脂酶Cβ3（PLCB3）在肝细胞癌（HCC）中的表达水平、预后评估价值及生物学功能。方法利用癌症基因组图谱数据库获取HCC相关样本mRNA测序数据，分析PLCB3在HCC组织和相邻正常组织中的表达差异。将上述样本中临床资料完整且存活时间≥30 d的样本分为PLCB3高表达组和PLCB3低表达组，分析PLCB3表达与两组患者总生存期、无进展生存期的关系，利用受试者工作特征曲线评估PLCB3表达对HCC患者预后的预测价值，比较PLCB3高、低表达组之间的临床病理特征、免疫细胞浸润情况，分析PLCB3表达与免疫检查点基因、特定药物半数抑制浓度（IC50）之间的关系。分析PLCB3高、低表达组之间的差异表达基因，并基于差异表达基因进行富集分析。从HCCDB v2.0网站下载单细胞数据集，进行单细胞分析和空间转录组分析。采用实时荧光定量PCR检测PLCB3在肝癌细胞和肝癌组织中的表达。结果 PLCB3在HCC组织中的表达水平高于相邻正常组织（P<0.05）；PLCB3低表达组患者的总生存期和无进展生存期均优于PLCB3高表达组（P<0.05），PLCB3表达水平对HCC患者短期生存率有较好的预测能力。在不同性别、肿瘤分级、肿瘤分期、肿瘤T分期、邻近肝脏炎症程度、胎蛋白水平的HCC患者中PLCB3的表达水平差异有统计学意义（P<0.05）。M2型巨噬细胞在PLCB3高表达组中显著富集，初始B细胞和Treg在PLCB3低表达组中显著富集；PLCB3表达与CD27、CD80等多个免疫检查点基因表达呈正相关，与CEACAM1、IL6R基因表达呈负相关（P<0.05）。PLCB3表达与达沙替尼等的IC50呈正相关，而与替吡法尼等的IC50呈负相关（P<0.05）。PLCB3与细胞外基质组织等生物学过程、离子通道复合体等细胞组分、单原子离子通道活性等分子功能密切相关，且主要参与神经活性配体-受体相互作用、脂肪酸代谢、过氧化物酶体等信号通路。单细胞分析显示，在肝细胞、肝星状细胞中，PLCB3在HCC组织中的表达水平高于相邻正常组织（P<0.05）；空间转录组分析显示，PLCB3主要分布于HCC肿瘤细胞、免疫细胞和基质细胞中。实时荧光定量PCR显示，PLCB3在Huh7、Hep3B和HCCLM3肝癌细胞系，以及HCC组织中高表达（P<0.05）。结论 PLCB3在HCC中高表达且与患者预后不良有关，其表达可能与细胞内信号传导通路的激活及肿瘤免疫微环境的改变有关，有可能成为潜在的预后标志物和治疗靶点。

Objective To investigate the expression, prognostic estimated value, and biological function of phospholipase Cβ3 (PLCB3) in hepatocellular carcinoma (HCC). Methods The mRNA sequencing data of HCC⁃related samples were obtained from the Cancer Genome Atlas database. The expression differences of PLCB3 in HCC tissues and adjacent normal tissues were analyzed. Samples with complete clinical data and survival time ≥30 days were divided into PLCB3 high⁃expression group or PLCB3 low⁃expression group. The relations between PLCB3 expression and the overall survival and progression⁃free survival of patients in the two groups was analyzed. The predictive value of PLCB3 expression for the prognosis of HCC patients was evaluated using the receiver operating characteristic curve. The clinical pathological characteristics and immune cell infiltration were compared between the PLCB high⁃expression and low⁃expression groups. The relations between PLCB3 expression and immune checkpoint genes, and specific drug half⁃maximal inhibitory concentration (IC50) was analyzed. The differentially expressed genes between the PLCB3 high⁃expression and low⁃expression groups were analyzed, and enrichment analysis was performed based on the differentially expressed genes. Single⁃cell data sets were downloaded from the HCCDB v2.0 website for single⁃cell analysis and spatial transcriptome analysis. Real⁃time fluorescence quantitative PCR was used to detect the expression of PLCB3 in hepatoma cells and liver cancer tissues. Results PLCB3 expression in HCC tissues was higher than that in adjacent normal tissues (P<0.05). The overall survival and progression⁃free survival of patients in the PLCB3 low⁃expression group were superior to those in the PLCB3 high⁃expression group (P<0.05). PLCB3 expression exhibited a good predictive ability for the short⁃term survival rate of HCC patients. There were statistically significant differences in the expression of PLCB3 between HCC patients with different gender, tumor grade, tumor stage, tumor T stage, adjacent liver inflammation degree, and fetoprotein level (P<0.05). M2⁃type macrophages were significantly enriched in the PLCB3 high⁃expression group, and initial B cells and Treg were significantly enriched in the PLCB3 low⁃expression group. PLCB3 expression positively correlated with the expressions of multiple immune checkpoint genes such as CD27 and CD80, and negatively correlated with the expressions of CEACAM1 and IL6R genes (P<0.05). The expression of PLCB3 positively correlated with the IC50 of dasatinib while negatively correlated with the IC50 of tipifarnib (P<0.05). PLCB3 was closely related to biological processes like extracellular matrix organization, cellular components including ion channel complexes, molecular function such as single⁃atom ion channel activity, and mainly participated in signaling pathways such as neural activity ligand⁃receptor interaction, fatty acid metabolism, and peroxisome. Single⁃cell analysis revealed that PLCB3 expression in HCC tissues was higher than that in adjacent normal tissue in hepatocytes and hepatic stellate cells (P<0.05). Spatial transcriptome analysis indicated that PLCB3 was mainly distributed in HCC tumor cells, immune cells, and stromal cells. Real⁃time fluorescence quantitative PCR interpreted that PLCB3 was highly expressed in Huh7, Hep3B, and HCCLM3 liver cancer cell lines, as well as in HCC tissues (P<0.05). Conclusion PLCB3 is highly expressed in HCC and is associated with poor prognosis in patients. Its expression may be related to the activation of intracellular signal transduction pathways and changes in the tumor immune microenvironment, and may become a potential prognostic marker and therapeutic target.

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