The Mechanism Of RFC5 In The Prognosis Of Cervical Cancer

Objective:To study the effect of replication factor C5(RFC5)on the proliferation and migration of cervical cancer Hela cells and the regulation of forkhead box M1(FOXM1)on RFC5,analyze the value of RFC5 in the diagnosis and prognosis of cervical cancer,establish a risk model of cervical cancer patients by using the immune genes related to RFC5,and establish a nomograph combining the risk model and clinical characteristics to evaluate the prognosis of cervical cancer patients.Methods:1.Construction of overexpression and interference with RFC5 expression lentivirus,construction of transcription factor expression vector,promoter luciferase vector and sea kidney luciferase reporter gene plasmid.2.After lentivirus infected Hela cells,the infection efficiency was detected by qRT-PCR and fluorescence microscope.3.The effects of RFC5 on the proliferation and migration of cervical cancer cells were analyzed by interference and overexpression of RFC5,CCK-8 and scratch test in Hela cells infected with lentivirus.4.The regulatory mechanism of transcription factor FOXM1 on RFC5 was analyzed by double luciferase report gene experiment.5.Download the transcriptome data and immunohistochemical results of cervical cancer in The Cancer Genome Atlas(TCGA),Gene Expression Omnibus(GEO)and Human Protein Atlas(HPA)databases,and analyze the expression of RFC5 in cervical cancer patients at the gene and protein levels respectively.6.Download the transcriptome data and clinical information of cervical cancer patients from the University of California Santa Cruz XENA(UCSC XENA)database,use R software to analyze the relationship between the expression of RFC5 and the prognosis of cervical cancer patients,and draw the receiver operating curve(ROC).7.Build the Protein–protein interaction(PPI)network diagram of the protein interacting with RFC5 in search tool for the retrieval of interacting genes/proteins(STRING)database,and conduct functional enrichment analysis.8.Analyze the immune genes related to RFC5 in TIMER2.0 database,and use R package to establish the risk scoring model of immune genes related to RFC5.Combined with the risk scoring model and clinical information of cervical cancer patients,a nomograph was constructed to evaluate the prognosis of cervical cancer patients.Results : 1.After lentivirus infects cells,qRT-PCR and fluorescence microscope confirm the infection efficiency.2.Compared with the negative control(NC)group,the overexpression of RFC5 promotes cell proliferation and migration,while the down-regulation of RFC5 inhibits cell proliferation and migration;3.FOXM1 combines with RFC5 promoter and luciferase activity increases.4.Biological information analysis showed that RFC5 was highly expressed in cervical cancer tissue,and the expression of RFC5 was significant correlated with the survival of cervical cancer patients.The area under curve(AUC)value of the ROC curve was 0.967,indicating that RFC5 was highly specific in the diagnosis of cervical cancer.5.The risk model of cervical cancer patients established with RFC5 related immune genes can be used as an independent prognostic factor for cervical cancer patients.The nomograph established by combining the risk scoring model with clinical information of cervical cancer patients can accurately predict the 3-year survival rate of cervical cancer patients.Conclusion : 1.RFC5 overexpression promotes the proliferation and migration of cervical cancer Hela cells.2.FOXM1 regulates the expression of RFC5 by binding with the promoter of RFC5.3.RFC5 can be used as a biomarker for diagnosis and prognosis of cervical cancer patients.4.The risk scoring model can be used as an independent risk factor for the prognosis of cervical cancer.The risk scoring model of cervical cancer patients and the nomograph established by clinical information can be used to evaluate the prognosis of cervical cancer patients.