Jiang Jinhua, Yan Tinghua, Guo Fang
Key Laboratory of Systems Biomedicine, Ministry of Education, Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, China.
Department of Interventional Oncology, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200127, China.
Curr Oncol. 2021 Sep 28;28(5):3758-3770. doi: 10.3390/curroncol28050321.
5-Hydroxymethylcytosine (5hmC) with dynamic existence possesses multiple regulatory functions. Whereas, 5hmC's impact on small hepatocellular carcinoma (SHCC) remains unclear. The present work focused on characterizing 5hmC content within SHCC and assessing the possibility of using global genomic 5hmC level as the predicative factor of clinical outcome.
This study applied ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) in measuring 5mC, 5fC and 5hmC contents. In addition, immunohistochemistry (IHC) was adopted to measure CK19 and 5hmC contents.
Research showed 5mC, 5hmC, and 5fC contents from global genomics of SHCC reduced extensively compared with healthy samples ( < 0.001). Moreover, SHCC was associated with lymph node metastasis (LNM). Greater 5mC and 5hmC levels were observed in non-metastasis group compared with the metastasis group ( < 0.001). Correlation analysis between the HBV DNA level and 5mC, 5fC and 5hmC levels exhibited that HBV DNA was associated with 5mC, 5hmC, and 5fC content reduction, which was verified in the cytological experiments. Moreover, 5hmC content had a negative correlation with the expression level of CK19 in SHCC. The decrease in 5hmC and CK19 containing 5hmC positive cell (called CK19) should be ascribed to the bad prognosis among SHCC patients.
The contents of 5hmC and CK19 of genomic DNA might be adopted for predicting SHCC survival as an important biomarker.
具有动态存在形式的5-羟甲基胞嘧啶（5hmC）具有多种调节功能。然而，5hmC对小肝细胞癌（SHCC）的影响仍不清楚。目前的工作重点是表征SHCC内的5hmC含量，并评估将全基因组5hmC水平用作临床结果预测因素的可能性。
本研究应用超高效液相色谱-串联质谱法（UHPLC-MS/MS）测量5mC、5fC和5hmC的含量。此外，采用免疫组织化学（IHC）法测量CK19和5hmC的含量。
研究表明，与健康样本相比，SHCC全基因组中的5mC、5hmC和5fC含量大幅降低（<0.001）。此外，SHCC与淋巴结转移（LNM）相关。与转移组相比，非转移组中观察到更高的5mC和5hmC水平（<0.001）。HBV DNA水平与5mC、5fC和5hmC水平之间的相关性分析表明，HBV DNA与5mC、5hmC和5fC含量降低有关，这在细胞学实验中得到了验证。此外，5hmC含量与SHCC中CK19的表达水平呈负相关。5hmC和含5hmC阳性细胞（称为CK19）的CK19降低应归因于SHCC患者的不良预后。
基因组DNA的5hmC和CK19含量可作为预测SHCC生存的重要生物标志物。
