目的: 探讨ANAPC4在肝细胞癌(hepatocellular carcinoma, HCC)中的表达水平及其临床意义,挖掘其潜在的作用机制。方法: 收集GEO、TCGA、SRA、GTEx、Oncomine及ArrayExpress数据库中HCC的mRNA表达数据集,整合分析ANAPC4的mRNA表达水平。通过THPA数据库下载ANAPC4免疫组织化学染色图片观察其蛋白表达强度。受试者工作特征曲线(receiver operating characteristic curve, ROC)、汇总受试者工作特征曲线、灵敏度、特异度、阴性和阳性似然比用于评估ANAPC4鉴别HCC的潜在价值。Begg检验用于评价数据发表偏倚。通过单细胞分析进一步验证ANAPC4的表达水平,CRISPR敲除筛选技术分析ANAPC4基因对于HCC细胞系生长的潜在影响。差异相关基因用于GO、KEGG通路富集分析。结果: 共35个数据集的样本(2 731 HCC组织vs 3 486正常组织)被纳入研究,合并标准化平均差(standardized mean difference, SMD)为0.65(95%CI:0.45-0.84),SROC曲线下面积为0.79(95%CI:0.75-0.83),灵敏度为0.62(95%CI:0.50-0.72),特异度为0.84(95%CI:0.74-0.90),阳性似然比为3.79(95%CI:2.50-5.75),阴性似然比为0.46(95%CI:0.36-0.58)。Begg检验提示不存在具有统计学意义的发表偏倚。分析单细胞测序数据提示ANAPC4在HCC细胞中表达显著上调, CRISPR-Cas9结果显示敲除ANAPC4显著抑制了HCC细胞的增殖能力。正相关高表达基因显著富集于细胞周期通路。结论: ANAPC4在HCC高表达,对HCC具有较好的鉴别能力,并可能通过细胞周期通路参与HCC的发生发展过程。
Objective: Comprehensive analysis of ANAPC4 expression and its clinical relevance in hepatocellular carcinoma (HCC), with exploration of underlying mechanisms. Methods: The mRNA expression data sets of HCC in GEO, TCGA, SRA, GTEx, Oncomine and ArrayExpress databases were collected, and the mRNA expression level of ANAPC4 was analyzed. The protein expression intensity of ANAPC4 was observed by downloading ANAPC4 immunohistochemical staining pictures from the THPA database. The receiver operating characteristic curve(ROC), summary receiver operating characteristics, sensitivity, specificity, negative and positive likelihood ratios were used to evaluate the potential value of ANAPC4 in the differential diagnosis of HCC. Begg test was used to evaluate data publication bias. The expression level of ANAPC4 was further verified by single cell analysis, and the potential effect of ANAPC4 gene on the growth of HCC cell lines was analyzed by CRISPR knockout screening technology. Differentially related genes were used for GO and KEGG pathway enrichment analysis. Results: A total of 35 data sets (2 731 HCC tissues vs. 3 486 normal tissues) were included in the study. The pooled standardized mean difference (SMD) was 0.65(95%CI: 0.45-0.84), the area under the SROC curve was 0.79(95%CI: 0.75-0.83), the sensitivity was 0.62(95%CI: 0.50-0.72), and the specificity was 0.84(95%CI: 0.74-0.90). The positive likelihood ratio was 3.79(95%CI: 2.50-5.75), and the negative likelihood ratio was 0.46(95%CI: 0.36-0.58). Begg test suggested that there was no statistically significant publication bias. Single-cell sequencing data analysis revealed that ANAPC4 is significantly upregulated in HCC cells, which was further confirmed at the single-cell level. CRISPR-Cas9 knockout experiments demonstrated that ANAPC4 deletion markedly impaired the proliferative capacity of HCC cells. Notably, genes positively correlated with high ANAPC4 expression were significantly enriched in the cell cycle pathway. Conclusion: ANAPC4 is highly expressed in HCC and has a good ability to identify HCC, and may be involved in the occurrence and development of HCC through the cell cycle pathway.
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