Wnt Pathway Mediated The Regulation Of Core Fucosylation Of Epithelial Cell By Lactobacillus Spp.in The Prevention And Treatment Of Cervical Cancer

Cervical cancer(CC)is the fourth most common malignant tumor in women.The morbidity and mortality are increasing year by year.Active and effective health interventions are urgently needed.Meanwhile,it is also important to analyze the pathogenesis of cervical cancer.Vaginal microecological imbalance often occurs in patients with cervical cancer,and fucosylation of mucosal epithelial cells is closely related to microbial colonization.Studies have found that compared with healthy women,fucosylation level in patients with cervical cancer is significantly lower.Core fucosyltransferase(Fut8)is the main glycosyltransferase that modifies protein fucosylation,and it plays an important role in the occurrence and development of multiple tumors.However,there is no report on the relationship between vaginal microbiota disorder and abnormal fucosylation of mucosal epithelial cells and its role in the occurrence and development of cervical cancer.Objective: Use gene knockout cell lines and animal models to deeply explore the internal relationship between vaginal microbiota disorder and abnormal protein fucosylation and its role in the prevention and treatment of cervical cancer.Methods:1、Using Lens Culinaris Agglutinin(LCA)to detect core fucosylation levels in serum,cervical exfoliated cells,cervical tissue and other clinical samples of healthy volunteers and cervical cancer patients;2、Constructing knockout cell lines and knockout mice,and explore the effects of core fucosyltransferase gene knockout on the proliferation and migration of cervical cancer cells;3、16S rRNA high-throughput sequencing analysis compares the composition of the vaginal microbiota of healthy volunteers and patients with cervical cancer,and explores the impact of HPV infection on the composition of the vaginal microbiota;4、MTT and scratch test to analyze the effect of Lactobacillus spp.on the proliferation and migration of cervical cancer cells;5、Analyze the correlation and underlying mechanism between Lactobacillus spp.and core fucosylation by Western bolt(WB)and chromatin immunoprecipitation-q PCR(CHIP-q PCR);6、Using WB and cellular immunofluorescence technology to analyze the effect of Lactobacillus spp.metabolite lactic acid on the core fucosylation modification of the vaginal mucosal epithelial cell and its internal mechanism.Results:1、Compared with healthy women,the level of core fucosylation in clinical samples such as cervical cancer patient serum,cervical exfoliated cells and cervical tissue was significantly reduced;2、After knocking out the Fut8 gene,the ability of cervical cancer cells to proliferate and migrate significantly increased;3、In patients with cervical cancer,the vaginal microbiota was disordered,the abundance of Lactobacillus spp.was significantly reduced,and the abundance of pathogenic bacteria such as Gardnerella spp.and Staphylococcus spp.was significant increased;HPV infection had little effect on the composition of vaginal microbiota;4、Lactobacillus iners,the dominant bacteria in the vagina of healthy women,significantly inhibits the proliferation and migration of cervical cancer cells;5、The abundance of L.iners was positively correlated with the level of core fucosylation;L.iners metabolites can promote the core fucosylation modification of epidermal cell by activating the Wnt pathway;6、L.iners metabolite lactic acid(Lactate)activates the Wnt pathway through Lactate-Gpr81 to up-regulate the core fucosylation level of vaginal mucosal epithelial cells.Conclusion:1、The level of core fucosylation in patients with cervical cancer was significantly reduced,and Fut8 gene knockout significantly can promoted the proliferation and migration of cervical cancer cells,which indicates that the fucosylation modification of epidermal cells had a protective effect;2、In patients with cervical cancer,the abundance of Lactobacillus spp.,especially L.iners,was significantly reduced;3、Lactic acid,a metabolite of L.iners,activated the Wnt pathway through the Lactate-Gpr81 method to up-regulate the core fucosylation level of vaginal mucosal epithelial cells,thereby inhibiting the proliferation and migration of cervical cancer cells,which regulates the vaginal microecology,and had an application prospects in cervical cancer prevention and treatment.