Center of Excellence in Molecular Genetics of Cancer and Human Diseases, Department of Anatomy, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
Tokyo Medical and Dental University, Tokyo, Japan.
Sci Rep. 2021 Jul 21;11(1):14838. doi: 10.1038/s41598-021-94330-1.
Early detection improves survival and increases curative probability in hepatocellular carcinoma (HCC). Peripheral blood mononuclear cells (PBMCs) can provide an inexpensive, less-invasive and highly accurate method. The objective of this study is to find the potential marker for HCC screening, utilizing gene expression of the PBMCs. Data from the NCBI GEO database of gene expression in HCC patients and healthy donor's PBMCs was collected. As a result, GSE 49515 and GSE 58208 were found. Using both, a statistical significance test was conducted in each gene expression of each data set which resulted in 187 genes. We randomized three selected genes (FLNA, CAP1, and CLU) from the significant p-value group (p-values < 0.001). Then, a total of 76 healthy donors, 153 HCC, 20 hepatic fibrosis, 20 non-alcoholic fatty liver were collected. Quantitative RT-PCR (qRT-PCR) was performed in cDNA from all blood samples from the qRT-PCR, The Cycle threshold (Ct) value of FLNA, CLU, CAP1 of HCC group (28.47 ± 4.43, 28.01 ± 3.75, 29.64 ± 3.90) were lower than healthy group (34.23 ± 3.54, 32.90 ± 4.15, 32.18 ± 5.02) (p-values < 0.0001). The accuracy, sensitivity and specificity of these genes as a screening tool were: FLNA (80.8%, 88.0%, 65.8%), CLU (63.4%, 93.3%, 31.3%), CAP1 (67.2%, 83.3%, 39.1%). The tests were performed in two and three gene combinations. Results demonstrated high accuracy of 86.2%, sensitivity of 85% and specificity of 88.4% in the FLNA and CLU combination. Furthermore, after analyzed using hepatic fibrosis and non-alcoholic fatty liver as a control, the FLNA and CLU combination is shown to have accuracy of 76.9%, sensitivity of 77.6% and specificity of 75%. Also, we founded that our gene combination performs better than the current gold standard for HCC screening. We concluded that FLNA and CLU combination have high potential for being HCC novel markers. Combined with current tumor markers, further research of the gene's expression might help identify more potential markers and improve diagnosis methods.
早期检测可提高肝细胞癌 (HCC) 的生存率和治愈率。外周血单核细胞 (PBMC) 可提供一种廉价、微创且高度准确的方法。本研究旨在利用 PBMC 的基因表达寻找 HCC 筛查的潜在标志物。
从 NCBI GEO 数据库中收集了 HCC 患者和健康供体 PBMC 基因表达的数据。结果发现 GSE49515 和 GSE58208。对每个数据集的每个基因表达进行统计意义检验，共得到 187 个基因。我们从有统计学意义的 p 值组（p 值<0.001）中随机选择三个基因（FLNA、CAP1 和 CLU）。然后，从所有血液样本的 cDNA 中进行了总共 76 名健康供体、153 名 HCC、20 名肝纤维化和 20 名非酒精性脂肪肝的定量 RT-PCR（qRT-PCR）。HCC 组的 FLNA、CLU 和 CAP1 的循环阈值 (Ct) 值（28.47±4.43、28.01±3.75 和 29.64±3.90）低于健康组（34.23±3.54、32.90±4.15 和 32.18±5.02）（p 值<0.0001）。这些基因作为筛查工具的准确性、灵敏度和特异性为：FLNA（80.8%、88.0%、65.8%）、CLU（63.4%、93.3%、31.3%）、CAP1（67.2%、83.3%、39.1%）。在两项和三项基因组合中进行了测试。结果表明，FLNA 和 CLU 组合的准确率为 86.2%，灵敏度为 85%，特异性为 88.4%。此外，在分析了肝纤维化和非酒精性脂肪肝作为对照后，FLNA 和 CLU 组合的准确率为 76.9%，灵敏度为 77.6%，特异性为 75%。此外，我们发现我们的基因组合的性能优于当前 HCC 筛查的金标准。我们得出结论，FLNA 和 CLU 组合具有成为 HCC 新型标志物的巨大潜力。结合当前的肿瘤标志物，进一步研究基因的表达可能有助于发现更多潜在的标志物并改善诊断方法。
