DataSheet1_Nanosecond pulse effectively ablated hepatocellular carcinoma with alterations in the gut microbiome and serum metabolites.PDF

Background: Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death in the world. Nanosecond pulsed electric fields (nsPEFs) have emerged as a new treatment for cancer. This study aims to identify the effectiveness of nsPEFs in the treatment of HCC and analyze the alterations in the gut microbiome and serum metabonomics after ablation.
Methods: C57BL/6 mice were randomly divided into three groups: healthy control mice (n = 10), HCC mice (n = 10), and nsPEF-treated HCC mice (n = 23). Hep1-6 cell lines were used to establish the HCC model in situ. Histopathological staining was performed on tumor tissues. The gut microbiome was analyzed by 16S rRNA sequencing. Serum metabolites were analyzed by liquid chromatography–mass spectrometry (LC-MS) metabolomic analysis. Spearman’s correlation analysis was carried out to analyze the correlation between the gut microbiome and serum metabonomics.
Results: The fluorescence image showed that nsPEFs were significantly effective. Histopathological staining identified nuclear pyknosis and cell necrosis in the nsPEF group. The expression of CD34, PCNA, and VEGF decreased significantly in the nsPEF group. Compared with normal mice, the gut microbiome diversity of HCC mice was increased. Eight genera including Alistipes and Muribaculaceae were enriched in the HCC group. Inversely, these genera decreased in the nsPEF group. LC-MS analysis confirmed that there were significant differences in serum metabolism among the three groups. Correlation analysis showed crucial relationships between the gut microbiome and serum metabolites that are involved in nsPEF ablation of HCC.
Conclusion: As a new minimally invasive treatment for tumor ablation, nsPEFs have an excellent ablation effect. The alterations in the gut microbiome and serum metabolites may participate in the prognosis of HCC ablation.

背景：肝细胞癌（Hepatocellular carcinoma, HCC）是全球范围内第三大癌症相关死亡病因。纳秒脉冲电场（Nanosecond pulsed electric fields, nsPEFs）作为一种新型癌症治疗手段逐渐得到应用。本研究旨在明确nsPEFs治疗肝细胞癌的疗效，并分析消融术后肠道菌群与血清代谢组的变化。
方法：将C57BL/6小鼠随机分为三组：健康对照组小鼠（n=10）、肝细胞癌模型组小鼠（n=10）以及经nsPEFs治疗的肝细胞癌模型组小鼠（n=23）。采用Hep1-6细胞系构建原位肝细胞癌模型。对肿瘤组织开展组织病理学染色。通过16S rRNA测序分析肠道菌群组成。采用液相色谱-质谱联用（liquid chromatography–mass spectrometry, LC-MS）代谢组学技术分析血清代谢物。通过斯皮尔曼相关分析探究肠道菌群与血清代谢组之间的关联。
结果：荧光成像结果显示nsPEFs具有显著的治疗效果。组织病理学染色观察到nsPEFs治疗组出现细胞核固缩与细胞坏死现象。nsPEFs治疗组中CD34、PCNA及VEGF的表达水平显著降低。与健康小鼠相比，肝细胞癌模型小鼠的肠道菌群多样性升高。Alistipes、Muribaculaceae等8个菌属在肝细胞癌模型组中富集，而在nsPEFs治疗组中这些菌属的丰度显著下降。LC-MS分析证实三组小鼠的血清代谢物存在显著差异。相关性分析表明，肠道菌群与参与nsPEFs消融肝细胞癌过程的血清代谢物之间存在关键关联。
结论：作为一种新型微创肿瘤消融治疗手段，nsPEFs具有优异的消融效果。肠道菌群与血清代谢组的变化可能参与肝细胞癌消融术后的预后调控。