| Objective The incidence of cervical cancer ranks second in female reproductive system tumors in China.At present,cervical cancer screening methods based on HPV genotyping combined with TCT often produce a large number of unnecessary colposcopy referrals or over-therapeutics,or have a certain rate of missed diagnosis because of their methodological limitations.Therefore,it is necessary to find specific molecular markers in cervical cancer to supplement the shortcomings of current early screening methods for cervical cancer.Foreign scholars have found that in HPV positive female patients,the degree of FAM19A4methylation increases with the severity of cervical lesions,and can be detected in almost all cervical cancers.It is proposed that FAM19A4 methylation is expected to be used as an alternative biomarker for early detection of cervical cancer,but there are few domestic studies on the diagnostic value of FAM19A4 gene methylation in cervical cancer.Based on the infection status of hr HPV gene subtypes in Inner Mongolia and China,this study explored the significance of FAM19A4 methylation alone or in combination with TCT or hr HPV genotyping in the diagnosis and treatment of cervical cancer and precancerous lesions in hr HPV positive women,in order to provide a basis for optimizing the existing cervical cancer screening and diagnosis strategies.Methods A total of 908 cervical exfoliated cells samples from patients with high-risk HPV positive(including 18 high-risk types:HPV16,18,31,33,35,39,45,51,52,53,56,58,59,66,68,73,82,83),which were collected in Inner Mongolia Cancer Hospital from February2018 to December 2021.All samples were strictly stored in the refrigerator at-80 degrees Celsius.After consulting the medical records,221 samples with complete clinical data,definite TCT or histopathological diagnosis and valid experimental results were selected for statistical analysis.The samples included normal cervix/cervicitis group(n=57),precancerous lesion group(n=119),and cervical cancer group(n=45).In the precancerous lesion group,there were 51 cases of CIN1,30 cases of CIN2 and 38 cases of CIN3.The following studies were carried out:1.To compare the distribution of different subtypes of hr HPV infection.2.To compare the difference of FAM19A4 methylation among normal cervical group,precancerous lesion group(CIN1,CIN2,CIN3)and cervical cancer group,and to analyze whether there is a trend between FAM19A4 methylation and the degree of cervical lesion.3.To compare the difference of FAM19A4 methylation detection rate among different hr HPV gene subtypes and different clinicopathological features(age,pathological type,FIGO stage,tumor size,lymph node metastasis).4.To explore the relationship between FAM19A4 methylation level and hr HPV,especially hr HPV16,52,58 and 18 infection.5.The receiver operating characteristic(ROC)curve was used to analyze the diagnostic efficacy of different detection methods,that is,FAM19A4 methylation,hr HPV16,52,58 and 18,cytology alone or in combination,in the diagnosis of CIN3 and above lesions,including sensitivity,specificity,PPV,NPV,accuracy and the corresponding 95%confidence interval,so as to comprehensively evaluate the diagnostic value of FAM19A4 methylation detection in hr HPV-positive women with cervical cancer and precancerous lesions.Results 1.Among the 908 hr HPV positive patients,the single infection rate of hr HPV was71.70%(651/908),and the multiple infection rate was 28.30%(257/908).The top four single hr HPV infections were hr HPV16,hr HPV52,hr HPV58 and hr HPV53,with detection rates of 18.92%,15.85%,12.15%and 8.15%,respectively.The detection rate of hr HPV16,52,58 combined with other hr HPV accounted for 57.58%of hr HPV multiple infection.2.The detection rates of FAM19A4 methylation in normal cervicitis/cervicitis group,CIN1,CIN2,CIN3 and cervical cancer groups were 17.5%(10/57),35.3%(18/51),36.7%(11/30),60.5%(23/38)and 82.2%(37/45),respectively.There was significant difference among the groups(P<0.05).In addition,there was a linear trend between different degrees of cervical lesions and FAM19A4 methylation(χ~2=46.845,P<0.05),that is,the detection rate of FAM19A4 methylation increased gradually with the increase of the severity of cervical lesions.The OR values of FAM19A4 methylation detection rate in CIN1,CIN3 and cervical cancer groups were 2.564(95%CI:1.051～6.255),7.207(95%CI:2.807～18.502)and 21.737(95%CI:7.802～60.563)respectively,which were significantly different from those in normal cervical/cervicitis group.3.There was no significant difference in the detection rate of FAM19A4 methylation among different age,histological type,clinical stage(FIGO,2018),tumor size and lymph node metastasis(P>0.05).4.In the cervical exfoliated cells of 221 cases of hr HPV positive patients included in this study,there was significant difference in the detection rate of FAM19A4 methylation among HPV16/18 positive patients,HPV52/58 positive patients and 14 other high risk types positive patients(P<0.05).By Spearman correlation analysis,it was found that there was a weak correlation(r=-0.155,P<0.05)between different HPV genotypes and the detection rate of FAM19A4 methylation.After the study population was divided into normal cervix group,precancerous lesion group and cervical cancer group,there was no significant difference in the detection rate of FAM19A4 methylation among different hr HPV positive patients(P>0.05).5.The sensitivity and specificity of FAM19A4 methylation,hr HPV16/18,hr HPV16/18/52/58 and TCT in≥CIN3 were 72.29%,71.74%,66.27%,74.64%,78.31%,47.83%,68.85%,70.63%,respectively.It is known that the sensitivity of FAM19A4methylation detection is between TCT and hr HPV16/18/52/58,the specificity is between TCT and hr HPV16/18,but the accuracy of FAM19A4 methylation is the highest.It’s AUC is the largest.In addition,although the sensitivity of FAM19A4 methylation was lower than that of hr HPV16/18 combined with TCT(72.29%vs 82.28%)or HPV16/18/52/58 combined with TCT(72.29%vs 88.61%),however,the specificity,PPV,accuracy and AUC were the highest,the value was 71.74%,60.61%,71.95%and 0.720,respectively.It shows that the detection of FAM19A4 methylation has good diagnostic value when it is used alone in≥CIN3.When FAM19A4 methylation was combined with hr HPV16/18,hr HPV16/18/52/58 and TCT,the sensitivity and NPV increased and the specificity decreased.The sensitivity and NPV of combination with hr HPV16/18 and TCT for≥CIN3 disease were 92.68%and 89.29%.The sensitivity of combination with hr HPV16/18/52/58 and TCT for≥CIN3 lesion was as high as95.12%and NPV was 88.24%,suggesting that it can be used alone or as a supplementary method for early screening of cervical cancer.The negative results of FAM19A4 methylation may be used as a reference index for excluding≥CIN3 lesions.Conclusion FAM19A4 methylation may be a specific biomarker to identify cervical cancer and precancerous lesions.The sensitivity of its combination with HPV16/18/52/58 and TCT to lesions above CIN3 is as high as 95.12%.It can be used alone or in combination with hr HPV typing test or TCT test for cervical screening and shunt in patients with positive hr HPV.It may be of value as a reference index for excluding lesions above CIN3. |
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