BackgroundPromoter methylation of MGMT and C13ORF18 has been confirmed as a potential biomarker for early diagnosis of cervical cancer. The aim of this study was to evaluate the performance of MGMT and C13ORF18 promoter methylation for triage of cytology screening samples and explore the potential mechanism.

MethodsMethylation-sensitive high-resolution melting was used to detect promoter methylation of MGMT and C13ORF18 in 124 cervical samples. High-risk human papillomavirus (HR-HPV) was detected by the Digene Hybrid Capture 2. Gene methylation frequencies in relation to cervical intraepithelial neoplasia (CIN) were analyzed. Frequencies were compared by Chi-square tests. The expression of gene biomarkers and methylation regulators was analyzed by immunohistochemical staining, real-time fluorescence quantitative polymerase chain reaction, and Western blot.

ResultsFor triage of low-grade squamous intraepithelial lesion (LSIL), gene methylation increased specificity from 4.0% of HR-HPV detection to 30.8% of MGMT (χ = 9.873, P = 0.002) and to 50.0% of C13ORF18 (χ = 21.814, P = 0.001). For triage of atypical squamous cells of undetermined significance, HR-HPV detection had higher positive predictive value of 54.8%. Either MGMT or C13ORF18 methylation combined with HR-HPV increased the negative predictive value to 100.0% (χ = 9.757, P = 0.002). There was no relationship between MGMT and C13ORF18 expression and DNA methylation (χ = 0.776, P = 0.379 and χ = 1.411, P = 0.235, respectively). MBD2 protein level in cervical cancer was relatively lower than normal cervical tissue (t = 4.11, P = 0.006).

ConclusionsHR-HPV detection is the cornerstone for triage setting of CIN. Promoter methylation of MGMT and C13ORF18 plays a limited role in triage of LSIL. Promoter methylation of both genes may not be the causes of gene silence.

Adult ; Cervical Intraepithelial Neoplasia ; genetics ; pathology ; Chi-Square Distribution ; DNA Methylation ; genetics ; DNA Modification Methylases ; genetics ; DNA Repair Enzymes ; genetics ; Female ; Humans ; Middle Aged ; Promoter Regions, Genetic ; genetics ; Squamous Intraepithelial Lesions of the Cervix ; genetics ; pathology ; Tumor Suppressor Proteins ; genetics ; Uterine Cervical Neoplasms ; genetics ; pathology ; Young Adult

OBJECTIVE: This study aimed to determine the factors affecting pathologic discrepancy and final diagnosis between colposcopic biopsy and pathology by loop electrosurgical excision procedure (LEEP). METHODS: Between 2004 and 2016, 1,200 patients who underwent LEEP were enrolled for this study. 667 underwent cervical cytology, human papillomavirus (HPV) test, colposcopic biopsy, and LEEP. We analyzed patient's age, menopausal status, number of delivery, abortion times, cervical cytology, number of punch biopsies, HPV type, LEEP, and interval between colposcopic biopsy and LEEP. RESULTS: Logistic regression analysis of the final diagnosis showed that age 30–39 years and other high HPV group types were associated with cancer diagnosis, whereas atypical squamous cells cannot exclude high-grade squamous intraepithelial lesion (ASC-H), high-grade squamous intraepithelial lesion (HSIL), and HPV type 16 affected the diagnosis of cervical intraepithelial neoplasia (CIN) 2. The overall concordance rate of histopathology between punch biopsy and LEEP was 43.3%. The rates of detecting a more severe lesion by LEEP than those by biopsy were 23.1%. The rates of a less severe lesion detected by LEEP than those by biopsy were 33.6%. Factors related with biopsy underestimation were as follows: < 1 vaginal delivery, HSIL, number of punch biopsies and HPV type. Punch biopsy number is a unique factor of biopsy overestimation. CONCLUSION: Patients with ASC-H, HSIL, and HPV type 16 may undergo conization immediately without colposcopic biopsy. We suggest that colposcopically directed 3 to 5 punch biopsies may be used to determine the need for conization.
Atypical Squamous Cells of the Cervix ; Biopsy* ; Cervical Intraepithelial Neoplasia ; Cervix Uteri* ; Conization* ; Diagnosis ; Female ; Humans ; Logistic Models ; Papanicolaou Test ; Pathology ; Squamous Intraepithelial Lesions of the Cervix

OBJECTIVE: To determine whether triage for atypical squamous cells of undetermined significance (ASC-US) and low-grade squamous intraepithelial lesion (LSIL) from the updated American Society for Colposcopy and Cervical Pathology cervical cancer screening guidelines is applicable in Korean women. METHODS: We investigated women with ASC-US or LSIL including referred from local hospitals visited for cervical cancer screening at Korea University Guro Hospital from February 2004 to December 2014. Detailed information on the results of Papanicolaou (Pap) smears, human papillomavirus (HPV) DNA tests, and cervical biopsies were collected through chart review. Cervical biopsy results were compared in eligible women according to individual Pap smear findings and HPV DNA status. RESULTS: Of 216,723 possible cases, 3,196 were included. There were 212 (6.6%) women with ASC-US and 500 (15.6%) with LSIL. The risk of ≥cervical intraepithelial neoplasia (CIN) 2 was significantly higher in women who were ASC-US/HPV+ than ASC-US/HPV- and LSIL/HPV+ than LSIL/HPV- (93.3% vs. 6.7% and 96.7% vs. 3.3%, P<0.001 and P<0.001, respectively). The risk of ≥CIN 3 was also significantly higher in women who were ASC-US/HPV+ than ASC-US/HPV- and LSIL/HPV+ than LSIL/HPV- (97.0% vs. 3.0% and 93.0% vs. 7.0%, P<0.001 and P<0.001, respectively). Age-stratified analysis revealed that more CIN 2 or CIN 3 was diagnosed in women aged 30 to 70 with ASC-US or LSIL when HPV DNA was present. CONCLUSION: Observation with Pap and HPV DNA tests rather than immediate colposcopy is a reasonable strategy for ASC-US or LSIL when the HPV DNA test is negative, especially in women aged 30 to 70. Reflection of these results should be considered in future Korean screening guidelines.
Atypical Squamous Cells of the Cervix* ; Biopsy ; Cervical Intraepithelial Neoplasia ; Colposcopy* ; DNA ; Female ; Human Papillomavirus DNA Tests ; Humans ; Korea ; Mass Screening* ; Papanicolaou Test ; Papillomaviridae ; Pathology* ; Squamous Intraepithelial Lesions of the Cervix* ; Triage* ; Uterine Cervical Neoplasms

OBJECTIVE: To identify key factors for predicting positive cone margin and appropriate cone length. METHODS: We retrospectively reviewed the margin status of patients who received conization with high grade cervical intraepithelial neoplasia, along with other factors such as patient age, parity, preoperative cytology, size of disease, type of transformation zone, and cone length from patient records. Cut-off value of cone length was analyzed in women younger than 40 years old because we design conization with minimum length especially for women who wish for future pregnancy. Cut-off value of cone length was defined as length corresponds to estimated probability of positive cone margin equal to 0.1 by logistic regression analysis with variables selected by stepwise methods. RESULTS: Among 300 patients, 75 patients had positive cone margin. Multivariable analysis revealed that squamous cell carcinoma at preoperative cytology (p=0.001), 2 or more quadrant disease (p=0.011), and shorter cone length (p<0.001) were risk factors for positive cone margin. Stepwise methods identified cone length and size of lesion as important variables. With this condition, cut-off value of cone length was estimated as 15 mm in single quadrant disease and 20 mm in 2 or more quadrant disease, respectively. CONCLUSION: We identified the independent risk factors of positive cone margin and identified the cut-off value of cone length to avoid positive cone margin in women younger than 40 years old. Conization should be performed not only according to colposcopic findings including type of transformation zone but size of disease and cone length.
Adult ; Cervical Intraepithelial Neoplasia/*pathology ; Cervix Uteri/*pathology ; *Conization ; Female ; Humans ; Middle Aged ; Retrospective Studies ; Uterine Cervical Neoplasms/*pathology

OBJECTIVE: We conducted a pooled analysis of published studies to compare the performance of human papillomavirus (HPV) testing and cytology in detecting residual or recurrent diseases after treatment for cervical intraepithelial neoplasia grade 2 or 3 (CIN 2/3). METHODS: Source articles presenting data on posttreatment HPV testing were identified from the National Library of Medicine (PubMed) database. We included 5,319 cases from 33 articles published between 1996 and 2013. RESULTS: The pooled sensitivity of high-risk HPV testing (0.92; 95% confidence interval [CI], 0.90 to 0.94) for detecting posttreatment CIN 2 or worse (CIN 2+) was much higher than that of cytology (0.76; 95% CI, 0.71 to 0.80). Co-testing of HPV testing and cytology maximized the sensitivity (0.93; 95% CI, 0.87 to 0.96), while HPV genotyping (detection of the same genotype between pre- and posttreatments) did not improve the sensitivity (0.89; 95% CI, 0.82 to 0.94) compared with high-risk HPV testing alone. The specificity of high-risk HPV testing (0.83; 95% CI, 0.82 to 0.84) was similar to that of cytology (0.85; 95% CI, 0.84 to 0.87) and HPV genotyping (0.83; 95% CI, 0.81 to 0.85), while co-testing had reduced specificity (0.76; 95% CI, 0.75 to 0.78). For women with positive surgical margins, high-risk HPV testing provided remarkable risk discrimination between test-positives and test-negatives (absolute risk of residual CIN 2+ 74.4% [95% CI, 64.0 to 82.6] vs. 0.8% [95% CI, 0.15 to 4.6]; p<0.001). CONCLUSION: Our findings recommend the addition of high-risk HPV testing, either alone or in conjunction with cytology, to posttreatment surveillance strategies. HPV testing can identify populations at greatest risk of posttreatment CIN 2+ lesions, especially among women with positive section margins.
Cervical Intraepithelial Neoplasia/pathology/surgery/*virology ; Female ; Humans ; Neoplasm Recurrence, Local/*virology ; Neoplasm, Residual ; Papillomaviridae/*isolation & purification ; Papillomavirus Infections/complications/*diagnosis ; Predictive Value of Tests ; Risk Assessment/methods ; Sensitivity and Specificity ; Uterine Cervical Neoplasms/pathology/surgery/*virology

OBJECTIVETo explore the expression of Snail and Slug in primary cervical squamous cell carcinoma (CSCC) and their relationship with KAI1 expression.

METHODSThe expressions of Snail, Slug, and KAI1 proteins were examined by immunohistochemistry in 154 specimens of CSCC tissues, 50 specimens of cervical intraepithelial neoplasm (CIN), and 40 specimens of normal cervical tissues.

RESULTSThe positivity rates of Snail, Slug, and KAI1 expression were 0%, 2.5%, and 95.0% in normal cervical tissues, 32.0%, 34.0% and 64.0% in CIN tissues, and 66.2%, 66.9%, and 43.5% in CSCC tissues, respectively, showing significant differences in the rates among the 3 groups (P<0.05). The expressions of Snail, Slug, and KAI1 were significantly correlated with the histological grades of the tumor, depth of invasion, lymph node metastasis, International Federation of Gynecology and Obstetrics (FIGO) stages, and postoperative survival time (P<0.05). The expressions of Snail and Slug were positively correlated (r=0.752, P<0.001), and both of them were negatively correlated with the expression of KAI1 (P<0.001). Kaplan-Meier analysis showed that patients positive for Snail and Slug had significantly lower survival rates than the negative patients (P<0.001), while a positive expression of KAI1 was associated with a higher survival rate of the patients. Cox regression analysis identified Snail, KAI1, and FIGO stage as independent factors that affected the outcomes of CSCC (P<0.05).

CONCLUSIONThe expressions of Snail, Slug, and KAI1 are related to the tumor grade, FIGO stage, invasive depth, lymph node metastasis, and prognosis of CSCC, and their combined detection can help estimate the outcomes of the patients.

Carcinoma, Squamous Cell ; metabolism ; pathology ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Kangai-1 Protein ; metabolism ; Kaplan-Meier Estimate ; Lymphatic Metastasis ; Neoplasm Staging ; Prognosis ; Snail Family Transcription Factors ; Survival Rate ; Transcription Factors ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; pathology

OBJECTIVETo investigate the differential expressions of nucleolin in invasive cervical squamous cell carcinoma, cervical intraepithelial neoplasms (CIN) and normal cervical epithelial tissues and explore the role of nucleolin in the carcinogenesis and progression of cervical squamous cell carcinoma.

METHODSFifty specimens of invasive cervical squamous cell carcinoma, 65 specimens of CIN, and 60 adjacent normal cervical epithelial tissue specimens were examined immunohistochemically for nucleolin expression. The correlation of nucleolin expression levels with histological grades of invasive cervical squamous cell carcinoma and CIN were analyzed.

RESULTSThe specimens of invasive cervical squamous cell carcinoma showed a significantly higher positivity rate for nucleolin expression than CIN and normal cervical epithelial tissues, and the rate in CIN tissues was significantly higher than that in normal cervical epithelial tissues (P<0.01). The expression level of nucleolin was significantly higher in invasive cervical squamous cell carcinoma than in CIN and normal cervical epithelia tissues, and higher in CIN than in normal cervical epithelia tissues, whose immunostaining scores were 7.6±0.3, 6.1±0.2, and 3.0±0.2, respectively (P<0.01). The mean nucleolin immunostaining score was significantly higher in poorly and moderately differentiated than in highly differentiated cervical squamous cell carcinoma (7.9 vs 7.1, P<0.01), and higher in high grade CIN than in low grade CIN tissues (6.0 vs 4.0, P<0.01).

CONCLUSIONSOverexpression of nucleolin plays an important role during carcinogenesis of cervical squamous cell carcinoma and is positively correlated with tumor progression of CIN and cervical squamous cell carcinoma.

Carcinogenesis ; Carcinoma in Situ ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; Disease Progression ; Female ; Humans ; Phosphoproteins ; metabolism ; RNA-Binding Proteins ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; pathology

OBJECTIVETo investigate the clinicopathological significance of Twist and YB-1 up-regulation in cervical cancer, and to correlate the expression of the two genes with E-cadherin, a marker of epithelial-mesenchymal transition (EMT).

METHODSA total of 202 tissue samples were collected during January 2008 to December 2013, including 50 cases of normal cervical tissues, 100 cases of cervical intraepithelial neoplasia (CIN) and 52 cases of squamous cell carcinoma (SCC). Twist, YB-1 and E-cadherin expression was investigated by MaxVision.

RESULTSIncreased expression levels of Twist and YB-1 were found and correlated with the malignant transformation of cervical epithelium, histological progression and metastasis of cervical cancer. In addition, Twist and YB-1 overexpression was also associated with aberrant expression of E-cadherin. Regression analysis revealed that Twist expression was an independent factor for the histological progression of cervical cancer.

CONCLUSIONSIt is suggested that Twist and YB-1 overexpression is significantly linked to cervical cancer tumorigenesis and progression, likely related to EMT through (YB-1)-Twist-(E-cadherin) pathway. Twist and YB-1 may be markers for determining the metastatic potential of cervical cancer.

Biomarkers, Tumor ; genetics ; metabolism ; Cadherins ; genetics ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Transformation, Neoplastic ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; Disease Progression ; Epithelial-Mesenchymal Transition ; Epithelium ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Nuclear Proteins ; genetics ; metabolism ; Twist-Related Protein 1 ; genetics ; metabolism ; Up-Regulation ; Uterine Cervical Neoplasms ; metabolism ; pathology ; Y-Box-Binding Protein 1 ; genetics ; metabolism

OBJECTIVETo investigate the expression of SOX2 in cervical intraepithelial neoplasia (CIN) and cervical cancer and explore its association with the clinical features.

METHODSSOX2 expressions were examined using immunohistochemical method in 10 normal cervical tissue specimens, 36 cervical intraepithelial neoplasia specimens (including 10 cases of grade I, 12 of grade II, and 14 grade III) and 40 cervical cancer specimens (including 21 cases of stage I and 19 of stage II). The correlation between the immunohistochemical results and the clinical features of the patients was analyzed.

RESULTSSOX2 expression was negative in normal cervical tissues, and was positive in 41.6% of CIN specimens (10.0% in CIN I, 41.7% in CIN II, and 64.3% in CIN III) in 82.5% of cervical cancer specimens (78.2% in stage I and 88.2% in stage II). The patients with cervical cancer had a significantly higher positivity rate of SOX2 than normal control group (P<0.05). The positivity rate of SOX2 increased with the evolution of cervical disease. SOX2 protein expression was significantly correlated with the histological grade and lymph node metastasis (P<0.05), but not with the age or clinical stage of the patients (P<0.05).

CONCLUSIONSOX2 expression may serve as a useful indicator for evaluating metastasis and malignancy of cervical cancer.

Cervical Intraepithelial Neoplasia ; genetics ; metabolism ; pathology ; Humans ; Lymphatic Metastasis ; Neoplasm Grading ; Neoplasm Staging ; SOXB1 Transcription Factors ; genetics ; metabolism

OBJECTIVETo evaluate the value of high risk (HR)-HPV viral load in predicting cervical lesions and triaging for HR-HPV positive women.

METHODSThe study cohort came from a multicenter cervical cancer screening program. HR-HPV was detected by hybrid capture 2 (HC-2) assay, and viral load was measured by the ratio of relative light units to cut off (RLU/CO). Women were divided into 4 groups according to the RLU/CO value, and CIN diagnostic system was used to describe the severity of cervical lesions. Chi-square trend test was used to analyze the association between viral load and CIN. The absolute and relative risks of CIN2+ in different viral load groups were calculated, and the clinical performance to detect CIN2+ at follow-up by different cut-off values of baseline RLU/CO was also calculated.

RESULTS2 725 women with complete results of both baseline and follow-up were included in this analysis. The severity of cervical lesions increased with the increasing viral load (P < 0.001). In women with normal or CIN1 diagnosis at baseline, the absolute risk of one-year accumulative CIN2+ was 0.11% in the HR-HPV-negative group, compared with 3.14% in the moderate viral load group and 6.09% in the high viral load group, and the relative risk of 29.05 (95%CI: 6.07-138.99) in the moderate viral load group and 56.34 (95%CI: 12.89-246.30) in the high viral load group. Raising cut-off value of baseline HR-HPV viral load to 15.00, RLU/CO decreased the number of women who need to be followed up at one-year from 774 to 412, with the sensitivity of 91.30% and specificity of 47.94% in detecting CIN2+ at follow-up.

CONCLUSIONSThe risk of cervical cancer and precancerous lesions increases with the increasing HR-HPV viral load. Raising the cut-off value of HR-HPV viral load can triage for HR-HPV-positive women, therefore help to allocate the health resources more effectively.

Adult ; Aged ; Cervical Intraepithelial Neoplasia ; pathology ; virology ; Disease Progression ; Female ; Follow-Up Studies ; Humans ; Mass Screening ; methods ; Middle Aged ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; Risk Factors ; Uterine Cervical Neoplasms ; pathology ; virology ; Viral Load