Infection Subtype And Site Of High-Risk HPV In Low-Grade Squamous Epithelial Lesions Of The Cervix

Background and objectiveCervical cancer(CC)is the third malignant tumor which is prevalent among women all over the world.Cervical cancer accounts for 9.8 percent of all cancers.About 528,000 new cases are reported in the word each year,resulting in about 270,000deaths.High risk human papillomavirus(HR-HPV)infection is closely related to the development of cervical cancer and Precancerous lesions.It is the most important risk factor,especially HPV 16 and 18.Cervical precancerous lesions contain low-grade intraepithelial neoplasia(LSIL)and high-grade intraepithelial neoplasia(HSIL).Their occurrence and development are associated with HPV infection.Most scholars believe that different cervical lesions are related to different HPV subtypes.The main HPV subtypes of HSIL infection are HPV 16 and 18.LSIL is mainly caused by low-risk HPV(LR-HPV)infection,such as HPV6 and 11.However,in clinical practice,the proportion of LSIL infected with HR-HPV is high.In this study,we compared HPV subtypes in LSIL and HSIL through test HPV using in situ hybridization(ISH),in order to To observe the infection sites and methods of HPV in different lesions.And To explore the new relationship between LSIL and HR-HPV.Materials and Methods1 The object of study:From October 2015 to December 2016,we collected 370paraffin samples which was diagnosed as cervical squamous intraepithelial lesion(SIL)through pathological examination from the department of pathology of zhengzhou university people’s hospital.The specimens were sliced and hematoxylin-eosin(HE)staining.The paraffin section was observed by two experienced pathologists.If they disagree with each other,the chief physician observed the slides and gave the final diagnosis.These samples included 190 LSIL and 180 HSIL.We observed sections with double blind method.None of the female patients had cervical lesions treatment history and exclusion of cases with other gynecological diseases.The age of the patients ranged from 18 to 79 years,and the median age was 38.This experiment has been approved by the ethics committee of Henan Province People’s Hospitalpatient.Patients and their families signed the informed consent.2 Research methods:HR-HPV genotype of 370 cervical tissues embedded by paraffin were detected by the methods of real-time fluorescence quantitative PCR(Roche Cobas4800 detection system).The subtypes were tested by roche Cobas4800 detection system include HPV 16,18 and other 12 HR-HPV types(31,33,35,39,45,51,52,56,58,59,66 and 68).Then we used HPV in situ hybridization to test paraffin samples and observe where the positive cells were located.Statistical software SPSS17.0 was used to analyze all data in this experiment.We usedχ~2 test and Fisher exact test methods to analyse data,and P<0.05 was considered the difference was statistically significant.Results1 Test results of HR-HPV in LSIL and HSIL:In this study,HR-HPV DNA of 370cases of cervical tissue samples were detected.The total infection rate of HR-HPV was95.9%(355/370).In LSIL and HSIL,the infection rate of HR-HPV was 94.2%(179/190)、97.8%(176/180),The total positive rate of HR-HPV in the two groups was not statistically significant(P=0.113).In LSIL and HSIL,HPV 16/18 positive rates were 26.3%、56.7%,respectively.Other HR-HPV(except for HPV 16/18)positive rates were79.5%、51.7%,respectively.The differences were statistically significant(P<0.001).2 LSIL and HSIL’s HE dyeing characteristics:HE staining was observed under the microscope.The surface epithelial cells of LSIL were well-differentiated and the cytoplasm was abundant.Hollow cells were formed,and the surface nuclei were atypical.Lesions were concentrated in the epidermal surface1/3.HSIL showed that cell polarity disappeared.There were a large number of abnormal mitotic and immature cells.Mitotic figure was present near the basal cells,and Nuclear atypical distributed extensively.The lesion extended upward from the basal layer,but the lesion was mainly concentrated in the 2/3 layers of the epithelium.3 Detection results of HPV in situ hybridization in LSIL and HSIL:HPV positive particles were stained brown-yellow in the nucleus.In situ hybridization results showed:In 127 HR-HPV positive patients,the brown-yellow cells of the 96 patients were confined to the surface of the cervical squamous epithelium and 31 patients were in the middle layer of the pithelium,and no positive virus particles were seen in the basal layer.In HSIL,the positive cells were more widely distributed,and the color was lighter.But they were mainly located in the lower 2/3 of the epithelium.Conclusions1 The positive rate of HR-HPV in LSIL was as high as 94.2%.HR-HPV infection is related to LSIL,especially other HR-HPV(except for HPV 16/18).2 The HR-HPV positive cells in LSIL were mainly located in the surface of the squamous epithelium;In HSIL,the positive cells were widely distributed,but they were mainly located in the lower 2/3 of the squamous epithelium.Different infection sites of HR-HPV can form two different lesions,LSIL and HSIL.