| Background and ObjectiveCervical cancer is one of the most common malignant tumors in women.Due to the development of cervical cancer screening technology,especially the extensive clinical application of cervical liquid-based cytology in recent years,the incidence of cervical cancer is significantly reduced.The International Cancer Society’s TBS(the Bethesda system for reporting cervical cytology 2001)grading reporting system classifies atypical squamous cells as smears with abnormal cells,suggesting there may be a potential cervical intraepithelial neoplasia without obvious cytological abnormalities.In Atypical squamous cell(ASC)smears,pathologists need to clearly identify atypical squamous cells of undetermined significance(ASCUS)or atypical squamous cells of cannot exclude high-grade squamous intraepithelial lesions(ASC-H),Because of ASCUS and ASC-H are not clear diagnoses,further examination is required to determine whether a high-grade squamous intraepithelial leision(HSIL)exists or may be develop.High-risk human papillomavirus infection is the most important risk factor for cervical cancer.According to TBS recommendations,Hybrid Capture 2 HPV testinghas been widely used in the classification of atypical squamous cells of undetermined significance patients.The sensitivity of HPV testing is good,but the high prevalence of transient HPV infection limits the specificity of HPV testing.Therefore,tumor markers of high sensitivity and specificity are need to find,in order to identify the HSIL in Atypical squamous cell patients.In 2013,the American Colposcopy and Cervical Pathology issued the latest screening guidelines for cervical cancer,which recommended that the clinical treatment of ASC-H should be the first choice for referral colposcopy.According to statistics,the detection rate of lesions above CINII in ASCUS was 6.90% ~ 23.27%,and the detection rate of lesions above CINII in ASC-H was 30.23% ~ 45.27%.Under-diagnosed and excessive diagnosis is clinical problems,and how to stratify the management of ASC patients has become a difficult problem.With the development of epigenetics,many studies have confirmed that DNA aberrant methylation plays an important role in tumor formation and development.Currently,numerous studies have confirmed that DNA methylation can be used for early detection,screening,diagnosis,prognosis evaluation and treatment of cervical cancer.In recent years,studies have shown that methylation of host genes PAX1 and ZNF582 can be used as an auxiliary biomarker to evaluate the risk of CIN progression.This study was based on hospital,through detecting the methylation level of PAX1 and ZNF582 genes,and in order to provide a reliable theoretical basis for ASC diagnosis and treatment,and establishes a new ASC triage and diagnosis model.Materials and methods Methods1.265 cases of effective residual specimens with ASC results from the liquid-based cytology of cervical cancer screening in the department of Gynecology clinic,the second affiliated hospital of Zhengzhou university from July 2017 to December 2018,and the effective residual specimens from patients who had undergone colposcopy and biopsy.The pathological biopsy results and medical history data were recorded in detail.2.The methylation levels of PAX1 and ZNF582 genes in cervical exfoliated cells of ASC population were determined by bisulfite conversion and quantitative methylation specific PCR(QMSP).And high-risk HPV detection by second-generation hybrid capture detection technology.3.Pathological results were used as the gold standard for patient stratification,To explore the diagnostic efficacy of PAX1 and ZNF582 methylation in detecting CINII+ and CINIII+ in ASCUS or ASC-H population.According to the area under the receiver’s operating characteristic curve(ROC).Chi-square test was used to compare the diagnostic effects between groups.The statistical analysis software used MedCalcV15.2,SPSS21.0,the test level α= 0.05,P <0.05,the difference was statistically significant.Results 1.Exploration of pathological results and cutoff value of ASC population(1)Among the 203 ASCUS patients,there were 86 cases of inflammation,65 cases of CINI,30 cases of CINII,17 cases of CINIII,1 case of cervical squamous cell carcinoma in situ,3 cases of cervical invasive squamous cell carcinoma,1 case of cervical invasive adenocarcinoma,and among the 62 ASC-H patients,there were 30 cases of inflammation,2 cases of CINI,4 cases of CINII,18 cases of CINIII,and 8 cases of cervical invasive carcinoma.(2)The ROC curve was used to analyze the cutoff values of PAX1 and ZNF582 in ASC patients diagnosed with CINII+,which were 8.8 and 11.1.At this time,the sensitivity was 85.5%,78.31%,and the specificity was 87.91%,92.86%.2.Diagnostic efficacy of three different detection methods for detecting CINII+ and CINIII+ in ASC population(1)PAX1 was found to be the best biomarker in the ASC population for the detection of CINII+.The sensitivity of PAX1 and HPV assays was 85.54% and 93.98%,and the specificities were 87.91% and 59.89%,respectively.ZNF582 is the best biomarker for the detection of CINIII+,the sensitivity of ZNF582 and HPV detection is 93.88% and 100.00%,and the specificity is 91.67% and 52.78%,respectively.(2)ROC curve was used to compare the area under the curve of HPVDNA,PAX1 and ZNF582 in the diagnosis of CINII+ and CINIII+.In the diagnosis of CINII+,the maximum area under the curve of PAX1 was 0.891(95%CI =0.847-0.926).In the diagnosis of CINIII+,the maximum area under the curve of ZNF582 was 0.960(95%CI=0.929-0.980).3.Diagnostic efficacy of three different detection methods for detecting CINII+ and CINIII+ in ASCUS population(1)In the ASCUS population,PAX1 was found to be the best biomarker for the detection of CINII +,the sensitivity of PAX1 and HPV detection were 83.02% and 90.57%,and the specificities were 87.33% and 64.67%,respectively.ZNF582 is the best biomarker for the detection of CINIII +,the sensitivity of ZNF582 and HPV detection is was 91.30% and 100.00%,and the specificity is 95.00% and 56.67%,respectively.(2)The ROC curve was used to compare the area under the curve of CINII+ and CINIII+ by HPV DNA,PAX1 and ZNF582.In the diagnosis of CINII+,the maximum area under the ROC curve detected by PAX1 was 0.862(95%CI=0.806-0.906).In the diagnosis of CINIII+,the maximum area under the curve of ZNF582 was 0.955(95%CI =0.859-0.943).4.Diagnostic efficacy of three different detection methods for detecting CINII+ and CINIII+ in ASC-H population(1)In the ASC-H population,ZNF582 was found to be the best biomarker for the detection of CINII +,the sensitivity of ZNF582 and HPVDNA was 90.00% and 100.00%,and the specificity was 90.62% and 37.50%,respectively.ZNF582 was the best biomarker to detect CINIII +,the sensitivity of ZNF582 and HPVDNA was 96.15% and 100.00%,respectively,and the specificity was 94.44 % and 33.33%.(2)ROC curve was used to compare the area under the curve of HPVDNA,PAX1 and ZNF582 in the diagnosis of CINII+ and CINIII+.In the diagnosis of CINII+ and CINIII+,the maximum area under the curve of ZNF582 was 0.862(95%CI=0.857-0.987)and 0.963(95%CI=0.881-0.994),respectively.5.Combined diagnosis of the three detection methods in ASC populationThe maximum area under the ROC curve of ZNF582 and PAX1 combined detection and diagnosis CINII+ was 0.946(95%CI =0.919-0.973),while the maximum area under the ROC curve of HPV,PAX1 and ZNF582 combined detection and diagnosis CINIII+ was 0.977(95%CI =0.958-0.995).However,the maximum area under the ROC curve of ZNF582 and PAX1 combined detection and diagnosis CINII+ was 0.976(95% CI=0.956-0.995),and there was no significant difference between the combined detection results of HPV,PAX1 and ZNF582(P>0.05).Conclusions1.The pathological results of cervical biopsy in ASC population have a wide range.Relatively speaking,the risk of CINIII+ in ASCUS population have a low risk,while those with ASC-H had a high risk.2.PAX1 or ZNF582 detection can more accurately identify lesions above HSIL or HSIL in patients with ASCUS or ASC-H compared with HR-HPV assay,and it is expected to replace HPV DNA detection as an effective means of triagepatients with atypical squamous cells.3.PAX1 and ZNF582 combined diagnosis can more accurately identify CINII+ lesions in ASCUS or ASC-H population,and the diagnostic effect of methylation detection has been confirmed. |
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