Establishment Of A Multi-factor Early Warning Model For The Occurrence And Progression Of Squamous Intraepithelial Lesion And MicroRNA29b And CCND2 Regulate The Mechanism Of SIL Progression

Part 1 Bioinformatics Analysis of Occurrence and Progress of SILObjective To study the genes and pathways related to the occurrence and progression of squamous intraepithelial lesion(SIL).Method Genemaina was used to build the gene network and the core genes were found in the network.Then the core genes related pathways which were associated occurrence and progression of SIL were screened in Kyoto Encyclopedia of Genes and Genomes databases(KEGG)and choose the core genes located in the common pathways related to SIL.Results The candidate pathways were identified as HPV infection signal pathway and Hippo signal pathway.The candidate genes were TGFBR2,CSKN1A1,PRKCI,FOXO1,CTNNB1,CCND2,MUC2,TP73,CTBP2 and PIK3 CA.Conclusions HPV infection signal pathway and Hippo signal pathway are related to the occurrence and progression of SIL.TGFBR2,CSKN1A1,PRKCI,FOXO1,CTNNB1,CCND2,MUC2,TP73,CTBP2 and PIK3 CA are the core components of the gene network.Part 2 Establishment of Multi-factor Predictive Models for The Occurrence and Progression of Squamous intraepithelial lesion Objective To study the risk factors for the occurrence and progression of squamous intraepithelial lesion(SIL)and establish predictive models.Methods A total of 30 cases of cervical squamous cells cancer(SCC),52 cases of SIL and 38 cases of normal cervix were enrolled in this study,and the tissue specimens of these cases were collected.The real-time fluorescence quantitative polymerase chain reaction(RT-QPCR)was performed to verify the different expression in cervical squamous cells cancer,SIL and normal cervix tissues respectively.R language was used for predictive models establishment and SPSS was use to statistical analysis.Results The clinical factors of these cases were studied.Compared to SIL ca ses,the age of SCC groups was older,the number of parity was more,and the pe rcentage of LSIL+ diagnosed by TCT was higher.The expression of TGFBR2,CSKN1A1,PRKCI and CTBP2 was significantly higher in SCC groups.While c ompared to normal cervix tissue,the percentage of HPV positivity and LSIL+ di agnosed by TCT was significantly higher.FOXO1 expression significantly high er in SIL tissue,but TGFBR2 and CTBP2 expression was significantly lower in SIL tissue.The significant genes and clinical factors were included in the model s.The best predictive models of SIL progression and occurrence were TGFBR2+CSKN1A1+PRKCI+FOXO1+CTBP2+TCT+menopause+parity+age and CTB P2+FOXO1+HPV+TCT,respectively.Conclusions Premenopause,older,TCT result with LSILI +,high parity,high expression of TGFBR2,CSKN1A1,PRKCI,and CTBP2,and low expression of FOXO1 are warning of SIL progression.HPV infection,TCT results with LSILI +,high FOXO1 expression and low CTBP2 expression indicate SIL occurrence.Part 3 The relationship between miR-29b/CCND2 and the occurrence and progression of squamous intraepithelial lesion Objective To study the relationship between miR-29 b and CCND2 and the occurrence and progression of squamous intraepithelial lesion.Methods Collect 30 cases of normal cervical tissue,SIL tissue and SCC tissue.ECT1 / E6E7 cells were selected as SIL cells and siha cells as cervical squamous cell car SILoma cells.miR29 b down-regulation and negative control lentivirals were transfected to ECT1/E6E7 and miR29 b up-regulation and negative control lentivirals were transfected to siha respectively.RT-QPCR was used to verify the expression of miR29 b and CCND2 in tissues and cells.Results Age,HPV infection,results of TCT and menopause were significantly different among normal group,SIL group and SCC group.The expression of miR-29 b in SIL tissue was significantly higher than that of SCC,but there was no significant difference in expression between SIL tissue and normal tissue.There was no significant difference in the expression of CCND2 in normal cervical tissue,SIL tissue and SCC tissue.The expression of miR-29 b and CCND2 in ECT1 / E6E7 cells was significantly higher than that of siha cells.Conclusions The expression of miR29 b in SCC tissues and siha cells is significantly down-regulated.CCND2 expression in siha cells was significantly reduced.The regulation disorder of miR29 b and CCND2 may be the cause of SIL progression.