The Correlation Between Vaginal Microflora And The Incidence And Stage Of Cervical Cancer

Background and purpose:Cervical cancer is one of the four most common cancers in women,seriously threatening women’s health.Early detection,early diagnosis and early treatment are the key roles for cervical cancer.The "three-step cervical cancer screening program" is commonly used in clinical practice to improve the early diagnosis rate of cervical cancer.However,this screening program has some limitations,such as high screening cost,high technical requirements for testing physicians and medical institutions,and insufficient coverage.Therefore,further optimization of cervical cancer screening methods is of great significance.Vaginal microflora is closely related to female reproductive health and participates in many reproductive diseases.Therefore,this study proposed to analysis the vaginal flora composition of the HR-HPV infection,cervical lesions and cervical cancer patients,from the point of the vaginal flora and through 16 S rRNA sequencing and bioinformatics methods.And exploring the changes of flora and looking for specific bacteria in HPV infection-cervical lesions-cervical cancer process,which for providing a new method to improve the screening effect and prognosis of cervical cancer patients.Content and methods:1.Specimen collection: A total of 155 eligible females who visited the outpatient or inpatient Department of the First Affiliated Hospital of Nanchang University from September 2019 to September 2020 were selected.Vaginal secretions of participants were collected,and the results of basic patient information,HPV test,cervical cytology,colposcopy,and cervical cancer FIGO stage were recorded.2.Sample processing: Bacterial DNA was extracted from vaginal secretions and amplified by 16 S rRNA gene V4 region.The amplified products were sequenced using Illumina Hi Seq 2500 platform.3.Data analysis: QIIME analysis process was used to analyze according to the following groups,mainly including Alpha diversity analysis,Beta diversity analysis,LEfse analysis,evolutionary tree analysis and so on.(1)Analysis of the difference of vaginal microflora in women infected with different types of HR-HPV:(1)HPV negative group,(2)HPV16/18 positive group,(3)Other HR-HPV positive group.(2)Analysis of the difference of vaginal microflora in women with different degrees of cervical lesions:(1)Control group,(2)ASC-US group,(3)SIL group,(4)CCa group(3)Analysis of the difference of vaginal microflora in cervical cancer patients with different stages:(1)Control group,(2)stage I group,(3)stage II group,(4)Stage III group.Result:1.Analysis of vaginal microfiora of women infected with different types of HPV(1)α-diversity analysis showed that compared with the HPV negative group,the abundance and diversity of female vaginal flora in HPV16/18 positive groups and other HR-HPV positive groups reduced,but the difference was not statistically significant.β-diversity analysis revealed significant differences in vaginal microflora among the three groups.(2)The abundance of Lactobacillus in HPV-negative group,Other HR-HPV positive group and HPV16/18 positive group decreased gradually,while the abundance of Gardnerella and Prevotella increased gradually.(3)Lefsefe analysis showed that,compared with the HPV negative group,the HPV16/18 positive groups were mainly enriched in Peptostreptococcu,Bacteroides,granulicatella,Anaerococcus,Arcanobacterium.While the other HR-HPV positive group were mainly enriched in Campylobacter.2.Analysis of vaginal microflora of women with different degrees of cervical lesions(1)α-diversity analysis showed that,compared with the Control group,the Shannon index increased gradually in the ASC-US group,SIL group and CCa group.And the β-diversity between four groups were significantly different.(2)The abundance of Lactobacillus in ASC-US group,SIL group and CCa group gradually decreased,while the abundance of Bacteroidia and Prevotella gradually increased.(3)Lefsefe analysis showed that Neisseria,Enterococcus,Staphylococcus and Ralstonia were enriched in the Control group.Lactobacillus,Bifidobacterium,Haemophilus were enriched in the ASC-US group.And Veronella was enriched in the SIL group.In CCa group,Bacteroidia,Clostridia,Prevotella,Sneathia,Anaerococcus,Peptostreptococcus and Fusobacterium were enriched.3.Analysis of vaginal microflora in patients with cervical cancer at different stages(1)α-diversity showed that compared with the Control group,the abundance and diversity of vaginal microflora in the three groups of cervical cancer stage I,II and III increased gradually,and the difference of β-diversity was statistically significant.(2)The abundance of Lactobacillus in the Control group was the highest,followed by the stage I group,and significantly decreased in the stage II and III groups.The abundance of Anaerococcus and Clostridia increased gradually in the four groups.(3)Lefsefe analysis showed that,at the genus level,the Control group was mainly enriched with Lactobacillus.The cervical cancer stage I was mainly enriched in Arcanobacterium and Deinococcus.The cervical cancer stage II was mainly enriched in Prevotella and Gemella.And the cervical cancer stage III was mainly enriched in Clostridia and Fusobacterium.Conclution:1.HR-HPV poaitive women,ASC-US,SIL and cervical cancer patients vaginal flora present disorder,and the composition of the flora was different from the control group.2.The abundance of Lactobacillus in HPV16 and 18 positive groups was lower than that in other high-risk HPV-positive groups,while Gardnerella and Prevotella were higher than that in other high-risk HPV-positive groups.3.The abundance of Lactobacillales decreased and the abundance of Bacteroidia and Prevotella increased with the aggravation of cervical lesions.4.The abundance of Lactobacillales decreased and the abundance of Anaerococcus and Clostridia increased with the increase of cervical cancer stages.