Differential Expression Of Cell Cycle Factors Geminin, Cdt1, P16 And Ki67 In Cervical Intraepithelial Neoplasia

Part Ⅰ Expression of Geminin and Cdt1 in normal tissues,chornic cervicitis and cervical intraepithelial neoplasmObjective:We aimed to measure the mRNA levels of Geminin and Cdc10 dependent transcript 1(Cdt1)in normal tissues,chronic cervicitis and cervical intraepithelial neoplasm(CIN).Our study might provide new biomarkers for screening and predicting the progression of cervical diseases.Methods:A total of 295(the average age was 38±12.5)cases were selected including 30 normal tissues,55 chronic cervicitis,70 CINⅠ,70 CINⅡ and 70 CINⅢ.Patients who had a history of chemotherapy,radiotherapy or immunotherapy were excluded from our study.This study was approved by the Ethnics Committee of the First Affiliated Hospital of Nanjing Medical University.Informed written consent was obtained from all subjects.RT-PCR was used to assess the mRNA levels of Geminin and Cdtl in all recruited tissues.Results:The mRNA levels of Geminin in normal tissues,chronic cervicitis,CINⅠ,CINⅡ and CINⅢ were 0.56±0.00,0.48±0.10,2.23±0.23,3.48±0.54 and 5.07±0.28,respectively.The mRNA levels of Cdtl in normal tissues,chronic cervicitis,CINⅠ,CINⅡ and CINⅢ were 1.11±0,00,1.01±0.58,2.87±1.24,4.09±1.64 and 5.52±1.36,respectively.The expression of Geminin and Cdtl were both increased with the severity of cervical diseases(p<0.05).However,there were no difference of Geminin and Cdtl expression between normal tissues and chronic cervicitis groups(p>0.05).Conclusions:Geminin and Cdtl were aberrantly expressed in cervical diseases.Our study suggested that Geminin and Cdtl might be involved in the development of CIN progression.Thus,Geminin and Cdtl might serve as potential biomarkers in cervical diseases.However,further functional studies are necessary.Part Ⅱ Analysis of Geminin,P16 and Ki67 in normal tissues,chornic cervicitis and cervical intraepithelial neoplasm and their correlations with HPV infectionObjective:To investigate the protein levels of Geminin,P16 and Ki67 in in normal tissues,chronic cervicitis and cervical intraepithelial neoplasm(CIN).In addition,we analyzed their correlations between human papillomavirus(HPV)infection.Methods:A total of 295 cases were selected including 30 normal tissues,55 chronic cervicitis,70 CINⅠ,70 CINⅡ and 70 CINⅡ.Patients who had a history of chemotherapy,radiotherapy or immunotherapy were excluded from our study.This study was approved by the Ethnics Committee of the First Affiliated Hospital of Nanjing Medical University.Informed written consent was obtained from all subjects.Immunohistochemistry was used to assess the protein levels of Geminin,P16 and Ki67 in all subjects.Spearman ’ s rank correlation coefficients were analyzed to investigate the possible correlations between HPV status and Geminin/P16/Ki67 expression.Results:Geminin and P16 were negative in all normal and chronic cervicitis tissues.Geminin was expressed in 12.9%of CINⅡ,54.3%of CINⅡ and 92.9%of CINⅢ(p<0.05).P16 was demonstrated in 15.7%of CINⅡ,61.4%of CINⅡ and 87.1%of CINⅢ(p<0.05).Ki67 was expressed in 3.3%of normal tissues and 5.5%chronic cervicitis tissues.The corresponding Ki67 expression in CINⅠ,CINⅡ and CINⅢ was 35.7%,82.9%and 95.7%,respectively.The median value of HPV status in chronic cervicitis,CINⅠ,CINⅡ and CINⅢ was 53.2,118.2,354.6 and 489.7(p<0.05).Specificity and PPV of Geminin were higher than those of p16 and Ki67.In addition,there were correlations between Geminin/Ki67 expression and HPV status.Conclusions:The sensitivity of sensitivity of Geminin was lower than that of P16 and Ki67.The negative predict value and accuracy of Geminin were comparable with P16 and Ki67.Geminin had a high degree of sensitivity and positive predict value in differentiation between CINⅠ and CINⅡ/Ⅲ.In addition,there were correlations between Geminin/Ki67 and HPV status.However,P16 expression was not related with HPV.Our study demonstrated that Geminin might serve as a new biomarker for the diagnosis of cervical diseases.Part Ⅲ Clinicopathologic characteristic of hysterectomy in cervical intraepithelial neoplasmⅡ/Ⅲ:a retrospective studyObjective:The objective of the study was to identify clinical issues concerning the hysterectomy in CINⅡ/Ⅲ:residual lesions,correlation between residual lesions and positive excision margin after cervical conization and risk of developing vaginal intraepithelial neoplasia(VAIN).We aimed to assess the necessity of hysterectomy in patients with CINⅡ/Ⅲ and the clinical value of follow-up.Methods:A total of 368 subjects of CINⅡ/Ⅲ treated with hysterectomy were retrospectively reviewed.We analyzed the cytology,HPV status,colposcope,biopsy after colposcope or hysterectomy,excision margin after cervical conization and follow-up vaginal stump.Results:40.5%patients were diagnosed as atypical squamous cells of undetermined significance(ASCUS)of Thinprep cytologic testresul(TCT)results.Hybrid CaptureⅡ System(HC2)detected for high-risk HPV load showed no correlation with CINⅡ/Ⅲ.Also,there was a relationship between positive excision margin after cervical conization and residual lesions.Additionally,11.1%patients developed VAIN.Conclusions:Our study suggested that the necessity of hysterectomy in patients with cervical intraepithelial neoplasmⅡ/Ⅲ is needed further discussion.In addition,the longer follow-up vaginal stump is warranted.