| Objective:1.Through investigation and analysis of cervical HR-HPV detection and cervical lesion detection in Karamay Central Hospital from January 2012 to December2021,to understand the changes of cervical HR-HPV infection and cervical lesion detection,and further to guide the prevention of cervical cancer in the region.2.To further clarify the application value of HPV E6/E7 m RNA detection in cervical cancer screening by analyzing the diagnosis of cervical lesions in HR-HPV positive women.3.By screening and verifying DNA methylation biomarkers which were meaningful for cervical cancer screening in exfoliated cell samples of different cervical lesions,further seeking meaningful DNA methylation biomarkers for cervical cancer screening.Methods:1.To collect the relevant data of women who were screened for cervical cancer in Karamay Central Hospital from January 2012 to December 2021.All subjects received HR-HPV testing,and with pathological diagnosis as the diagnostic standard of cervical lesions.To understand the trend of HR-HPV infection and cervical lesions detection vertically,and understand the common subtypes of HR-HPV infection in this region horizontally.2.To collect relevant data on women who tested positive for HR-HPV DNA during cervical cancer screening,using pathological diagnosis as the"gold standard"for diagnosis,to understand the expression of HPV E6/E7m RNA in different lesions and the significance of diagnosing cervical HSIL+.And compare the application value of HPV E6/E7 m RNA detection and TCT in the diagnosis of cervical HSIL+.3.To collect cervical exfoliated cell samples with positive HR-HPV in cervical cancer screening,selecting 6 cases of LSIL-and 9 cases of HSIL+samples,to analyze DNA methylation status by Illumina850K methylation chip technology.To screen out meaningful methylation genes for pyrophosphate sequencing,and verify the diagnostic significance of HSIL+in DNA methylation markers.Results:1.From January 2012 to December 2021,the total number of women who underwent cervical HR-HPV screening in Karamay Central Hospital was 85429,of which the population proportion of women aged 36 to 45 was the largest,and which of women over 55 years old was the smallest.The overall infection rates of HR-HPV from2012 to 2021 were 16.40%(1376/8391),20.81%(1479/7106),25.20%(1472/5842),19.71%(1950/9895),13.80%(1263/9153),13.42%(1010/7528),12.98%(1021/7864),12.86%(1044/8119),11.47%(1153/10048),and 11.04%(1268/11483),respectively.The annual changes showed a downward trend(APC=-7.34%),and the differences were statistically significant(P<0.05).There was significant statistical difference in the infection rate of HR-HPV among different age groups from 2012 to 2021(χ~2=120.015,P<0.05),with the highest infection rate being observed in the age group over 55 years old.The infection rate of HR-HPV in all age groups showed downward trend from 2014 to 2021,and the differences in annual change trend were statistically significant(P<0.05).The overall detection rates of cervical HSIL+or LSIL+in HR-HPV infections from 2012 to 2021showed upward trend,and the differences in annual change trends were statistically significant(P<0.05).However,the detection rate of cervical cancer in cervical LSIL+shows a decreasing trend(APC=-12.65%,P<0.05).The detection rates of HSIL+or LSIL+in the age groups of 25-35 years old,36-45 years old,and 46-55 years old showed upward trend from 2012 to 2021,and the differences in annual change trends were statistically significant(P<0.05).The total detection rate of HSIL+or LSIL+in the age group over 55 years old was the highest among all age groups,with 18.66%(95/509)or23.97%(122/509).However,there was no significant upward trend in the annual detection rates of HSIL+or LSIL+in the age group over 55 years old.The common subtypes of HR-HPV in this region are HPV52(17.28%),HPV16(13.76%),HPV58(11.50%),HPV56(10.75%),HPV51(10.15%).The proportion of single infection in cervical HR-HPV was 80.20%(1316/1641),and the proportion of mixed infection was 19.80%(325/1641).2.The positive rate of HPV E6/E7m RNA in HSIL+was 77.85%(116/149),and in LSIL-was 27.27%(39/143),and there was statistically significant difference between the two(P<0.05).In the diagnosis of cervical lesions,the complete coincidence rate between HPV E6/E7m RNA diagnosis and pathological diagnosis was 75.34%(220/292),and the Kappa value of diagnostic consistency test was 0.506.There was no statistically significant difference in the diagnosis of cervical lesions in different types of HR-HPV DNA using HPV E6/E7m RNA(P>0.05).There was no significant statistical difference in the diagnosis of cervical lesions by HPV E6/E7 m RNA in different age groups(P>0.05).The positive expression of HPV E6/E7m RNA in different cervical lesion tissues increases with the severity of cervical lesion(χ~2=75.749,P<0.05).Moreover,the positive expression of HPV E6/E7m RNA was moderately positively correlated with the severity of the lesion(r=0.517,P<0.05).The sensitivity of HPV E6/E7m RNA in diagnosing HSIL+was 77.9%(95%CI:68.1%～78.2%),while the sensitivity of TCT in diagnosing HSIL+was 73.2%(95%CI:73.1%～82.6%),and there was no significant statistical difference between the two(P>0.05).The specificity of HPV E6/E7m RNA in diagnosing HSIL+was 72.7%(95%CI:68.1%～78.2%),while the specificity of TCT in diagnosing HSIL+was 53.1%(95%CI:47.4%～58.9%),with significant statistical difference between the two(P<0.05).The Youden index of HPV E6/E7 m RNA for diagnosing HSIL+was 0.506,higher than TCT(0.263).To calculate the AUC for diagnosing HSIL+,HPV E6/E7m RNA was 0.753,TCT was 0.632,“HPV E6/E7m RNA and TCT”was 0.705,and“HPV E6/E7m RNA or TCT”was 0.680.The consistency between HPV E6/E7m RNA and TCT diagnosis of HSIL+and LSIL-was poor.HPV E6/E7m RNA had no predictive significance on pathological changes after HSIL+cervical conization.There was no significant statistical difference in the diagnosis of cervical lesions between HPV E6/E7m RNA in cervical tissue and cervical exfoliated cell samples(χ~2=3.413,P>0.05).3.DNA methylation analysis was conducted on samples of different cervical lesions that passed DNA extraction and quality inspection.A total of751 DMP sites were selected,which were evenly distributed on each chromosome.Compared to the LSIL-group,the HSIL+group had 641 high methylation sites and 105low methylation sites.Two genes,GSTM5 and PRR23C,corresponding to cg25210835,cg24467349,and cg24530147,cg06508738,cg02930239,were screened out from them.Two sites of PRR23C,cg24530147 and cg06508738,were screened out through pyrophosphate sequencing.And expanded sample experiments were conducted on the two sites and five other detected sites.The results showed that the average methylation rate of the HSIL+group was higher than that of the LSIL-group in this region,Among them,the methylation rate of the cg24530147 site and the other two Cp G sites in the HSIL+group were significantly higher than that of the LSIL-group,and the differences were statistically significant(P<0.05).Conclusions:1.From 2012 to 2021,the overall infection rate of HR-HPV showed downward trend,while the detection rates of cervical HSIL+or LSIL+showed overall upward trend.However,the detection rate of cervical cancer in cervical LSIL+showed downward trend.The proportion of annual screening population for women over 55 years old was the smallest,but the HR-HPV infection rate,HSIL+or LSIL+detection rate were the highest among all age groups.The common subtypes of HR-HPV in this region were HPV52,HPV16,HPV58,HPV56,and HPV51.2.The positive rate of HPV E6/E7m RNA in HSIL+was significantly higher than that in LSIL-and the positive expression of HPV E6/E7m RNA showed upward trend with the severity of cervical lesions.The ability of HPV E6/E7m RNA to diagnose HSIL+was significantly superior to TCT,and it has good application value in cervical cancer screening 3.This experiment discovered a new DNA methylation gene PRR23C in cervical cancer screening,among which three loci showed a significantly higher methylation rate in the HSIL+group than in the LSIL-group,which had certain application prospects in cervical cancer screening. |
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