Correlation Studies Between HPV Persistent Infection, Cervical Cancer Occurrence And Development And HPV16L1and LCR Methylation In Uighur Women

Objective: To investigate type-specific persistent infection of HPV in Uighur womenand associated society/behavioral risk factors; to evaluate the relationship between HPV16L1,LCR methylation levels and HPV persistent infection, cervical cancer occurrence anddevelopment.Methods: HPV positive and random selective HPV negative women whoattended the cervical cancer screening women in September2012to September2013inthe Xinjiang Uighur autonomous region people’s hospital outpatient as screening objectswere done by questionnaire survey, and HPV positive women did HPV genotypes test andbiopsy under colposcope detecting pathological diagnosis, to follow-up the women ofchronic cervicitis for12months. Using Logistic regression to analyses its influencingfactors; Using pyrophosphate sequencing technology to detect positioning and quantitativemethylation levels of L1,LCR in HPV16positive groups of five pathological levels(persistent infection, transient infection, CIN1, CIN2-3, cervical cancer), Using ROCcurve analysis to evaluate the value of HPV16methylation to diagnose CIN2+and predictHPV persistent infection;Using Immune histochemical method to inspect the expression ofL1protein in the5pathological levels; Using multiple real-time PCR to detect HPV16E2/E6ratio to indicate the physical state. Results: Type-specific HPV persistence in Uighurwomen was25.5%,HPV16(48.00%), HPV18(31.03%), HPV58(28.30%), HPV52(23.40%),HPV45(21.43%)were the most frequently persistent types among major types. Viralload>1000pg/ml compared to viral load in1≤HPV≤100(OR:2.40,95%CI:1.11-5.15),HPV16infection compared to non-HPV16infection(OR:4.81,95%CI:2.63-8.18)increasedthe risk of type-specific HPV persistent infection; Multivariate analysis showed thatcompared to the traisend infection, postmenopause (OR:2.41,95%CI:1.23-2.89), not touse condoms (OR:3.85,95%CI:1.68-4.58) increased the risk of type-specific persistentHPV infection. Methylation rates of13sites in L1were statistically significant differencesamong5pathological lesions groups and as the disease progresses, methylation rates were on the rise; high methylation in13CpG sites of L1increased the risk of CIN2+, thehighest value of OR was9.89for site6650; AUCs of13CpG sites in L1for the diagnosisof CIN2+were between0.756to0.862, the highest value is site6650with the AUC of0.862; High methylation of site6389、6457、6581、6650、6796、7034increased the riskof persistent infection, P<0.05, the strongest risk estimate site was site6389(OR:13.33,95%CI:3.95-28.08)；AUCs of these sites in L1for the prediction of HPV persistentinfection were between0.656to0.943, the highest value is site6389with the AUC of0.943; The methylation rate of site31、37、43、52and58in LCR were statisticallysignificant differences among five level of pathological lesions and as the diseaseprogresses, methylation rates were on the rise; high methylation in these CpG sitesincreased the risk of CIN2+, the strongest risk estimate site was site58(OR:5.71,95%CI:2.54-12.84), AUCs of these CpG sites for the diagnosis of CIN2+were between0.640to0.848, the highest value of site is site58with the AUC of0.848; methylation rateof CpG sites in promoter, E2BS3and E2BS4was on the decline with increase ofproportion of integrated status in CIN2-3and cervical cancer, the difference wasstatistically significant, P<0.05. methylation rate of CpG sites in cervical cancer ofintegrated status were between11.50%to14.25%. Conclusion: Uighur women withHPV16positive, high viral load, menopause, not to use condom will likely increase riskfrom a transient into the persistent infection, Should strengthen follow-up of the crowd.Methylation rates of HPV16L1has high value to predict HPV persistent infection,cervical cancer lesion progress; The methylation of CpGs in the promoter region wascommon in HPV16LCR region, which indicated that the methylation of promoter is veryimportant in the regulation of transcription.