Research For The Application Of P16, Ki67 Semi-quantitative Score On The Diagnosis Of CIN And Its Relationship With HPV Infection

Objective:1.The combined application of P16, Ki-67 immunohistochemical staining will be used to make a classification diagnosis of cervical intraepithelial neoplasia( CIN),by the semi quantitative score.We will also explore its role in pathological diagnosis in CIN.2. To further clarify the relationship between the expression of P16 protein and HPV infection we observed the expressed situation of P16 and the distribution of infectioned HPV and its subtypes in different levels.of cervical lesions.Methods:Collecting 671 patients with the diagnosis of ASCS and above by TCT, in Outpatient Department of the second hospital of Shanxi Medical University during June to December of 2014, diagnosed them by HE staining pathological: 375 cases of chronic cervical inflammation, 269 CIN which included 140 CIN I,66 CIN II,63 CIN III.(included 45 CIN II~III, 18 CIN III),27 cases of cervical cancer. Divided into inflammation groups,CIN I,CIN II,CIN III, Squamous cell carcinomas.Results:IV 1. The incidence of P16 and KI- 67 positive rate were 38.30%, 37.85%, respectively, along with the rise of the cervical lesion level enhancement, both closely related(Spearman rank correlation coefficient = 0.931, 0.876, < 0.001). Set in different pathological changes, P16 positive rate were 0%, 72.14%, 100%, 100%, 100%; KI- 67 positive rate were 0%, 70%, 100%, 100%, 100%, the difference between groups were statistically significant(< 0.001); And in the second category of cervical CIN, P16, Ki67 positive expression differences between groups are statistically significant(< 0.001). Bothhigh sensitivity for the forecast of CIN lesions, strong specificity.2. Immunohistochemical semi-quantitative score results showed that: in inflammation group,P16 and Ki67 all express negative(100%). In CIN I group,mainly express: both expressed as 2 +(32.14%), Ki67 negative and P16 expressed as 2 +(25%), both expressed as 1 +(22.14%), P16 expressed as 1 +and KI-67 expressed as 2 +(10%); CIN II group main expression in the combination: P16 expressed as 2 +and KI-67 expressed as 3 +(56.06%), P16 expressed as 3 +and KI-67 expressed as 2 +(43.94%); Both in CIN III group expressed as 3 +(77.78%), the remaining(22. 22 %) at least one of the indicators can be judged for 4 +.3. The incidence combined P16 and Ki67 semi-quantitative score to diagnose CIN pathology classification, comparing to conventional HE pathological grading diagnosis research. Cervical chronic inflammation in the group diagnosis consistent rate was 100%. CIN I in accordance rate was 95%(133/140), 7 cases can not only make an accurate classification or classification do not agree with HE classification. Above CIN II grade, diagnostic consistency of 100%, according to the second category can be diagnosed with HSIL(129/129).4. The total incidence of HPV infection rate was 73.62%, different lesion group infection rates were 62.4%, 80%, 95.45%, 80%, 94.44% and 100%. Respectively,with cervical lesion level rise higher positive rate.In the HPV classification tests, the top six infected subtype are: type 16(26.97%), 58(6.40%), 18(4.02%), 52(3.43%), 39(2.98%),33(2.83%). Mixed infection is given priority to with high-risk type mixed a total of 110 cases, among them at high risk of mixed infection(15.50%). Statistical results show that the type HPV16 infection and cervical CIN lesions of secondary classification degree wasstatistically significant(P = 0.002)5.As the P16 expression intensity increasees, HPV infection rate increased accordingly. P16 expression of negative, + 1, + 2, + 3, + 4 infection rates were 62.08%, 84.78%, 92.31%, 94.19%, 97.05%. And HPV16, 58, 52, 31, 66 and in high-risk hybrid P16 negative and positive comparison difference was statistically significant(P < 0.05).Methods:1.All the cases which show cytology positive were detected HPV types and cervicalcolposcopy biopsy.2.Cervical biopsy specimens were embedded by paraffin HE staining.Three respectively pathologists made the pathological diagnosis and its classification. 3.The immunohistochemical staining of P16,KI-67 were given to the most serious lesions, Three respectively pathologists made semi-quantitative score.Then,a pathology classification diagnosis of cervical lesions would be made with thoes comprehensive analysis of the two semi-quantitative score.4. Statistical Methods:Using SPSS 19.0 statistical processing, with P < 0.05 for the difference was statistically significant. Meet the needs of normal distribution of quantitative data using statistical description, otherwise using classification data expressed in frequency; Meet the needs of normal distribution between multiple sets of quantitative data comparison with variance analysis, quantitative data can not meet the needs of normal distribution using nonparametric test, classifying data comparison between groups of using 2? inspection; Hierarchical data by Spearman rank correlation analysis.Conclusion:1.The semi-quantitative score of P16 and Ki- 67 immunohistochemical staining can reflect the degree of CIN lesions objectively.2.HE staining observed by Optical microscope,combined with the P16 and Ki- 67 immunohistochemical semi-quantitative score can improve the accuracy and repeatability of the pathological diagnosis of CIN, providing more credible therapeutic basis for clinic.3.Different subtypes of HPV infection show different performance in this cases,which is mainly for the 16 type(26.97%), and more 58 type(6.40%) than 18(4.02%), and should be focused on.4.The detection of P16 show a certain significance in the prognosis and outcome of cervical CIN lesions with HPV infection.