The Value Of P16/Ki67 Double Staining In The Diagnosis Of Cervical Squamous Intraepithelial Lesions

Objective:To collect 131 cases of cervical biopsy specimens from January 1,2016 to December 31,2017 in gynecological clinics and hospitalized patients at the Second Hospital of Jilin University,After the pathological specimens were again confirmed by HE staining and P16 and Ki67 immunohistochemical staining,the above pathological sections were re-cut for P16/Ki67 immunohistochemical double staining,and the expression of P16 and Ki67 in single staining and double staining was analyzed and compared.To explore the value of P16/Ki67 double staining in distinguishing cervical LSIL from HSIL.Methods:P16/Ki67 double staining was performed on the collected specimens using antiP16(H6E4)mouse monoclonal antibody,Ki67 antibody reagent,AP staining solution,etc.And under the microscope to observe the expression of cervical lesions in different degrees again.SPSS 21.0 statistical software was used for data analysis.The difference was considered statistically significant at P<0.05.Chi-square test was used for counting data,and the consistency of positive expression of P16 and Ki67 in single and double staining results was compared by Kappa analysis method.Results:A total of 131 cases of cervical lesions were stained in this study.Among them,27 cases were chronic cervicitis and 104 cases were SIL tissues(including 40 cases of LSIL and 64 cases of HSIL).In the inflammatory,LSIL,and HSIL sections,the positive expression rates of P16 were 3.70%,52.50%,and 84.38%,respectively(P<0.05).In the light microscopy of LSIL tissues,the expression of P16 was mostly scattered and patchy.In light microscopy of HSIL tissue,P16 is mostly full-scale,continuous expression.In the inflammation group,LSIL group,and HSIL group,the positive rates of Ki67 were 0.00%,60.00%,and 89.06%,respectively(P<0.05);in the light microscopy of LSIL,the expression of Ki67 is mostly located in the basal and parabasal layers.In the tissues of HSIL under light microscope,the expression of Ki67 mostly reached or exceeded 2/3 of the squamous epithelium or was highly expressed.For the P16/Ki67 double staining,the positive expression of P16 and Ki67 in the inflammation group,LSIL and HSIL groups were statistically significant(P<0.05).At the same time,under the light microscope,there was no significant difference in the coloring position and degree of staining between P16 and Ki67 in the cells,and there was no significant difference between the clarity and single staining.When the double staining was performed during the reading,the distribution and relationship of P16 and Ki67 in the tissue cells could be directly observed,which was more intuitive than the multiple stained sections.We analyzed the co-expression of P16 and Ki67 in double staining and showed the correlation between P16 and Ki67 double staining in different degrees of cervical squamous intraepithelial lesions.The data showed that during the increase of lesion levels,P16,The incidence of common positive Ki67 also increased.The positive expression of P16 and Ki67 was significantly different between the inflammation group,LSIL and HSIL groups(P<0.05).After statistical analysis,the positive expressions of P16 and Ki67 in single stain and double stain were respectively analyzed,and the two diagnostic methods were tested for Kappa consistency.In this study,positive expression of P16 in single stain and double stain was performed.The value of Kappa coefficient was 0.854,indicating that P16 has good agreement in immunohistochemistry for single staining and double staining.At the same time,the Kappa coefficient value of the positive expression of Ki67 in single stain and double stain was calculated to be 0.826,indicating that Ki67 has good consistency in single staining and double staining of immunohistochemistry.Conclusions:1.This study validated the significance of P16 and Ki67 in the diagnosis and differential diagnosis of inflammatory tissue,LSIL,and HSIL.Combined immunohistochemical staining with P16 and Ki67,HE-assisted HE staining can help pathologists more accurately diagnose SIL.2.The value of P16/Ki67 double staining in the diagnosis of SIL was demonstrated.3.Validated the value of P16/Ki67 double staining in inflamed tissues,LSIL,and HSIL.The positive expression of the two in diseased tissues reflects the severity of the lesions,which helps to identify LSIL and HSIL,and it can be certain.Degree to overcome the reality of clinical diagnosis.