Yang Chia-Yu, Yeh Yuan-Ming, Yu Hai-Ying, Chin Chia-Yin, Hsu Chia-Wei, Liu Hsuan, Huang Po-Jung, Hu Song-Nian, Liao Chun-Ta, Chang Kai-Ping, Chang Yu-Liang
Department of Microbiology and Immunology, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
Front Microbiol. 2018 May 3;9:862. doi: 10.3389/fmicb.2018.00862. eCollection 2018.
Oral squamous cell carcinoma (OSCC) is a highly aggressive cancer and the fourth leading malignancy among males in Taiwan. Some pathogenic bacteria are associated with periodontitis and oral cancer. However, the comprehensive profile of the oral microbiome during the cancer's progression from the early stage to the late stage is still unclear. We profiled the oral microbiota and identified bacteria biomarkers associated with OSCC. The microbiota of an oral rinse from 51 healthy individuals and 197 OSCC patients at different stages were investigated using 16S rRNA V3V4 amplicon sequencing, followed by bioinformatics and statistical analyses. The oral microbiota communities from stage 4 patients showed significantly higher complexity than those from healthy controls. The populations also dynamically changed with the cancer's progression from stage 1 to stage 4. The predominant phyla in the oral samples showed variation in the relative abundance of , and . The abundance of increased significantly with the progression of oral cancer from the healthy controls (2.98%) to OSCC stage 1 (4.35%) through stage 4 (7.92%). At the genus level, the abundance of increased, while the number of , and decreased with cancer progression. , and were associated with OSCC, and they progressively increased in abundance from stage 1 to stage 4. The abundances of , and were inversely associated with OSCC progression. We selected a bacterial marker panel of three bacteria (upregulated , down-regulated , and ), which had an AUC of 0.956 (95% CI = 0.925-0.986) in discriminating OSCC stage 4 from the healthy controls. Furthermore, the functional prediction of oral bacterial communities showed that genes involved in carbohydrate-related metabolism, such as methane metabolism, and energy-metabolism-related parameters, such as oxidative phosphorylation and carbon fixation in photosynthetic organisms, were enriched in late-stage OSCC, while those responsible for amino acid metabolism, such as folate biosynthesis and valine, leucine, and isoleucine biosynthesis, were significantly associated with the healthy controls. In conclusion, our results provided evidence of oral bacteria community changes during oral cancer progression and suggested the possibility of using bacteria as OSCC diagnostic markers.
口腔鳞状细胞癌（OSCC）是一种侵袭性很强的癌症，在台湾男性中是第四大常见恶性肿瘤。一些病原菌与牙周炎和口腔癌有关。然而，在癌症从早期发展到晚期的过程中，口腔微生物群的综合概况仍不清楚。我们对口腔微生物群进行了分析，并确定了与OSCC相关的细菌生物标志物。使用16S rRNA V3V4扩增子测序，随后进行生物信息学和统计分析，对51名健康个体和197名不同阶段的OSCC患者的口腔冲洗液中的微生物群进行了研究。4期患者的口腔微生物群群落比健康对照组的群落复杂性显著更高。随着癌症从1期发展到4期，菌群数量也动态变化。口腔样本中的主要门类在相对丰度上表现出变化。随着口腔癌从健康对照组（2.98%）发展到OSCC 1期（4.35%）再到4期（7.92%），其丰度显著增加。在属水平上，随着癌症进展，其丰度增加，而、和的数量减少。、和与OSCC相关，并且它们的丰度从1期到4期逐渐增加。、和的丰度与OSCC进展呈负相关。我们选择了由三种细菌组成的细菌标志物组合（上调的、下调的和），其在区分OSCC 4期和健康对照组时的曲线下面积（AUC）为0.956（95%置信区间=0.925 - 0.986）。此外，口腔细菌群落功能预测显示，参与碳水化合物相关代谢（如甲烷代谢）的基因以及与能量代谢相关的参数（如氧化磷酸化和光合生物中的碳固定）在OSCC晚期富集，而负责氨基酸代谢（如叶酸生物合成以及缬氨酸、亮氨酸和异亮氨酸生物合成）的基因与健康对照组显著相关。总之，我们的结果提供了口腔癌进展过程中口腔细菌群落变化的证据，并表明了将细菌用作OSCC诊断标志物的可能性。
