| Background and PurposeCervical Cancer(CC)is a common malignancy which threatens women's life and health.Its incidence and mortality rate rank second in women's three most common malignancies.The development of cervical cancer lesions,from low grade squamous intraepithelial neoplasia(CIN1)to invasive carcinoma(ICC),takes about ten years.Epidemiological and molecular biological studies indicate that the occurrence and development of cervical cancer are closely related to the persistent infection of high risk human papillomavirus(hrHPV).Reducing the incidence of uterine cervix cancer and the probability of death,so that patients receive timely and effective diagnosis and treatment,which is currently the focus of the study.Therefore,the more effective early screening measures for cervical cancer are of great significance.At present,cervical intraepithelial lesions are mainly detected by using TCT in combination with HPV DNA.Traditional detection of HPV DNA has certain advantages.It can distinguish between high-risk and low-risk HPV,but its detection has some limitations.The limitations are as follows:1.Traditional HPV DNA tests cannot distinguish among active,persistent,and progressive infections.2.HPV DNA can also express in apoptotic cells and cell debris,which result in some false positives.HPV genomics studies have shown that HPV coding includes 6 early open reading frames(E1,E2,E4,E5,E6 and E7)and 2 late reading frames(L1 and L2).HPV E6/E7 is a viral oncogene that significantly mediates the degradation of tumor suppressor genes p53 and Retinoblastoma protein(Rbp),thereby interfering with cell cycle regulation,which can cause malignant transformation of host cells.HPV E6/E7mRNA exhibits oncogene activity in cervical tissue.Previous literature studies have shown that HPV E6/E7 mRNA levels correlate with the severity of cervical intraepithelial lesions.Seterdal et al.and Molden et al.found that HPV mRNA has high specificity and positive predictive value compared with HPV DNA.Foreign studies have found that most human telomerase mRNA(hTERC)gene is abnormally amplified in cervical epithelial neoplasia,and the abnormal expression of this gene is cervical tumor.The basis of the occurrence may also be an early event of cervical cancer formation.Therefore,the hTERC gene test can not only judge the condition of the lesion,but also assess its development.One single indictator of HPV E6/E7 mRNA and hTERC in cervical intraepithelial lesions has been studied at home and abroad,however,the combined study on cervical lesions has not been reported in the literature.In summary,this study used branched-chain DNA technology(b-DNA)and fluorescence in situ hybridization(FISH)to detect HPV E6/E7 mRNA and hTERC in different degrees of cervical lesion histology and cells.The specificity,sensitivity,positive predictive value,and negative predictive value of the combined detection of the HPV E6/E7 mRNA and the hTERC gene were analyzed.This study was to investigate the clinical value of HPV E6/E7 mRNA and hTERC in the diagnosis and evaluation of cervical cancer risk progression in order to provide a basis for the early screening of cervical cancer.Materials and Methods1.Collect 135 samples of cervical fluid-based cytology from July 2016 to October 2017 in the Yellow River Central Hospital.According to the Bethesda system(TBS)of cervical cytology,the sample content was as follows:27 cases in NILM group,36 cases in ASCUS group,16 cases in LSIL group,13 cases in ASC-H group,7 cases in AGC group,30 cases in HSIL group and SCC group in 6 patients,the diagnosis is based on histopathology.The sample content was as follows:normal/inflammation in 27 cases,CIN1 in 35 cases,CIN2 in 33 cases,CIN3 in 28cases,and ICC in 12 cases.In this study,the final test results for all patients were histologically diagnosed as the gold standard.2.b-DNA was used to detect HPV E6/E7 mRNA levels in cervical fluid based cytology specimens.FISH was used to detect the hTERC gene amplification in cervical fluid-based cytology specimens.3.Compare HPV E6/E7 mRNA expression status and observation of hTERC gene amplification in different grade histology and different grade cytology specimens.4.SPSS 19.0 software was used for data statistics to analyze the specificity,sensitivity,positive predictive value and negative predictive value of a single index and the combination of the two.X~2 test was used among different groups,and the difference was statistically significant at P<0.05.Results1.The expression of HPV E6/E7 mRNA in different histological grades:In 135specimens,the positive rate of HPV E6/E7 mRNA was 65.19%(88/135).The positive expression rate of HPV E6/E7 mRNA in the normal/inflammation group,CIN1 group,CIN2 group,CIN3 group and ICC group increased sequentially,33.3%(9/27),45.7%(16/35),78.8%(26/33),89.3%(25/28)and 100%(12/12).The HPV E6/E7 mRNA expression level was significantly different in different histological grades of cervical cervix(?2=34.18,P<0.01).2.HPV E6/E7 mRNA expression in different grades of cytological classification:In 135 specimens,HPV E6/E7 mRNA in NILM group,ASCUS group,LSIL group,ASC-H group,AGC group,HSIL group and SCC group.The rate of positive expression in the middle of the population increased to 33.3%(9/27),52.8%(19/36),56.3%(9/16),84.6%(11/13),and 100%(7/7),90%(27/30),100%(6/6).The level of HPV E6/E7 mRNA expression was significantly different in different cytological diagnosis grades of cervical(?2=32.32,P<0.01).3.Amplification of hTERC gene in histological classification at different levels:In 135 specimens,the positive rate of hTERC gene amplification was 49.63%(67/135).The positive rate of hTERC gene amplification in the normal/inflammation group,CIN1 group,CIN2 group,CIN3 group and ICC group increased sequentially,7.4%(2/27),14.29%(5/35),63.6%(21/33),96.4%(27/28)and 100%(12/12).The difference in the level of hTERC gene amplification between different histological grades of the cervix was statistically significant(?2=76.04,P<0.01).4.Amplification of hTERC gene in different grades of cytological classification:In 135 specimens,hTERC gene was positively expressed in NILM group,ASCUS group,LSIL group,ASC-H group,AGC group,HSIL group and SCC group.The rates increased sequentially,7.4%(2/27),19.4%(7/36),56.3%(9/16),61.5%(8/13),85.7%(6/7),and 96.7%(29/30),100%(6/6).The level of hTERC gene amplification was significantly different in different cytological diagnosis grades of the cervix(?2=69.68,P<0.01).5.HPV E6/E7 mRNA,hTERC gene detection and the combined detection of the two methods were compared:the specificity,sensitivity and positive predictive value of the combined detection were higher compared with the HPV E6/E7 mRNA detection and the hTERC gene test.The specificity,sensitivity and positive predictive value among the three methods were statistically significant(?2,11.15,8.81,8.66,P<0.05).Conclusions1.The sensitivity,specificity and positive predictive rate of combined detection of HPV E6/E7 mRNA and hTERC gene were higher than those of the two methods alone.2.The expression of HPV E6/E7 mRNA and hTERC gene both reflect the development of cervical lesions.The combined detection of HPV E6/E7 mRNA and hTERC gene can increase the early screening rate of cervical cancer. |
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