Objective To determine the relationship between local microenvironment and cervical cancer (CC). Discovering differential florae and metabolites associating with CC. Design Observational study. Participants 10 LSIL patients, 10 HSIL patients, 10 CC patients and 10 healthy controls were enrolled in our study. Methods We performed 16S rDNA sequencing and metabolomic analysis in the cervicovaginal fluid to reveal the differential florae and metabolites during cervical carcinogenesis. Results Carcinogenesis was associated with alterations in microbiome diversity, individual taxa, and functions. Bacterial diversity was increased and the composition was changed by the influence of cervical cancer carcinogensis. Lactobacillus was significantly reduced in LSIL and CC patients. On the contrary, the relative abundance of Prevotella was significantly elevated in LSIL patients. Lipids were significantly declined in LSIL and HSIL patients compared to HC participants, and significantly elevated in CC compared to LSIL and HSIL patients. Pathway enrichment analysis found phenylalanine, tyrosine and tryptophan biosynthesis pathway was crucial in cervical carcinogenesis. Conclusions These results showed that microbic and metabolomic profiling are capable of distinguishing CC from precancer and highlighted potential biomarkers for the early detection of cervical dysplasia. These differential microorganisms and metabolites expected to become a potential tool to assist in the diagnosis of cervical cancer. Funding We would like to thank Development & Demonstration Program of Wuxi (N20192004), Key Research & Development Program of Jiangsu Province (BE2015617), The 5th Phase of “Project 333” of Jiangsu (BRA2019024) and Innovation and Entrepreneurship Training Program for College Students in Jiangsu Province (KYCX19_1182) for their support.

Cervical cancer (CC) continues to be one of the most common cancers among females worldwide. It takes a few years or even decades for CC to arise in a minority of women with cervical precancers. An increasing corpus of studies today indicates that local microecology and carcinogenesis are intimately related. To investigate the changes in cericovaginal microecology with the development of cervical cancer, we performed 16S rDNA sequencing and metabolomic analysis in cericovaginal fluid from 10 LSIL patients, 10 HSIL patients, 10 CC patients and 10 healthy controls to reveal the differential flora and metabolites during cervical carcinogenesis. Carcinogenesis is associated with alterations in microbiome diversity, individual taxa, and functions with notable changes in Lactobacillus, Prevotella and Aquabacterium, as well as in cervicovaginal metabolites that correlate with cervicovaginal microbial patterns. Increased bacterial diversity and a decline in the relative abundance of Lactobacillus, the dominant species in the cericovaginal flora, are observed when cervical lesions advance. According to KEGG pathway enrichment analysis, lipids and organic acids change as cervical cancer progresses, and the phenylalanine, tyrosine, and tryptophan biosynthesis pathway is essential for the development of cervical cancer. Our results reveal that microbic and metabolomic profiling is capable of distinguishing CC from precancer and highlights potential biomarkers for the early detection of cervical dysplasia. These differential microorganisms and metabolites are expected to become a potential tool to assist in the diagnosis of cervical cancer.

Cervical cancer(CC)continues to be the second leading cause of cancer death in women aged 20 to 39[1].Recent research has shown that the cervical microbiota differs in different stages of cervical carcinogenesis[2].It has been reported that the gut microbiota is linked to host metabolism,which influences the progression of the disease[3].The role of cervicovaginal flora in affecting host metabolism,thereby causing the progression of cervical cancer,requires further research.In this direction,metabolomics has been implemented to detect small molecular differential metabolites in cancer progression.Metabolomics is the study of the thousands of low-molecular-weight molecules found in biological fluids and tissues of different individuals,whether normal or afflicted with disease,and it reflects reasonable changes in biological functions.

Toll样受体(Toll Like Receptor,TLR)是一种模式识别受体( pattern recognition receptor , PRR) ,微生物及其代谢产物入侵人体是通过识别PRR,激活固有免疫从而激活抗原特异性获得性免疫系统[1-2].TLRs是一个膜受体家族,在感知包括细菌、真菌和病毒在内的多种入侵病原体方面发挥着关键作用[3-4].TLR4是人类发现的第一个TLR,是介导信号转导的重要PRR,而且是唯一可以经髓样分化因子88 ( myeloid differ-entiation primary response protein 88 , MyD88 )依赖型和β干扰素TIR结构域衔接蛋白( TIR domain-containing adaptor indu-cing interferon-β,TRIF)依赖型两条信号通路的TLR家族成员.TLR4具有多种不同结构的配体,如植物二萜紫杉醇、呼吸道合胞病毒的融合蛋白、纤连蛋白和热休克蛋白,其中最重要的配体是脂多糖[1].脂多糖是革兰阴性菌外膜的重要成分,有强大的免疫刺激活性,TLR4通过识别脂多糖激活下游信号通路,在机体的细菌感染和免疫反应中起着重要作用.