AIM2 Was Involved In Cervical Epithelial Lesions With HPV Infection

BackgroundCervical cancer(CCa)is a common malignant tumor of the female reproductive system.Cervical Squamous intraepithelial lesions(SIL)is cervical precancerous lesions which is divided into Low grade Squamous intraepithelial lesions(LSIL)and high-grade Squamous intraepithelial lesion(HSIL).LSIL corresponding CIN I of the three-level classification and HSIL corresponding CIN Ⅱ and Ⅲ of the three-level classification.The development from SIL to CCa is a long and complex process.Currently,High Risk Human Papillomavirus(HR-HPV)infection is known to be the primary Risk factor for the occurrence and development of cervical cancer and its precancerous lesions,but the specific mechanism of HR-HPV in cervical epithelial lesions has not been clearly determined,so further research is needed.It is worth noting that in recent years,the detection rate of cervical SIL is increasingly high and the age of onset is getting younger and younger.And cervical biopsy pathology diagnosis as diagnostic gold standard of SIL with strong subjectivity,has been facing the difficulty of diagnosis of CIN I.,II which lead to excessive clinical treatment.Therefore,the search for reliable molecular markers of cervical SIL is of great importance for adjuvant pathological diagnosis and grading,and then accurate treatment.Absent in Melanoma 2(AIM2),which is a double-stranded DNA(dsDNA)receptor in the cytoplasm,and is an important immune molecule of the body in response to exogenous dsDNA virus infection.AIM2 binds to the intracytoplasmic dsDNA and together with the adaptor protein ASC and caspase-1,forms the in:flammatory masome,and releases Il-18 and Il-1β,and induces Pyroptosis,which plays an important role in the innate immune response.Recent studies have shown that AIM2,in addition to its role in infectious diseases,may also affect the occurrence and development of a variety of tumors.Combined with SIL and CCa,viral infector-related diseases were defined.This study detected and discussed the expression of AIM2 in cervical epithelial lesions with HPV infection,its role in the occurrence and development of the disease,and its diagnostic value in cervical epithelial lesions.ObjectivesThis paper takes AIM2 as the entry point to define the expression characteristics of AIM2 in cervical lesions associated with HPV infection.And we will investigate the diagnostic value of AIM2 in diseases and the role of AIM2 in cervical benign epithelial cells with HPV infection.The research may provide a new perspective and a new theory for the in-depth understanding of the occurrence and development of cervicallesions with HPV infection.Methods1.Paraffin specimens of Normal cervical(HPV-),chronic cervicitis(HPV+),LISL,HSIL and cervical squamous cell carcinoma were collected and Pathological sections were made.Immunohistochemical staining of AIM2 was performed to compare the differences of the expression levels of AIM2 protein in cervical lesions of each group.HPV+/-Cervical cancer cell lines and cervical benign epithelial cell lines were collected and cultured,and the mRNA and protein expression levels of AIM2 in each cell line were detected by RT-qPGR and Western blot.2.The tissue microarray of cervical lesion tissue were made and the immunohistochemical staining of AIM2 and P16 was performed to compare the expression differences between AIM2 and P16.And the diagnostic value of the two indicators in distinguishing LSIL and HSIL was compared by ROC curve.3.AIM2 overexpression plasmid was designed and transiently transfected AIM2 plasmid into human cervical benign epithelial cell line H8.CCK-8 assay,Edu proliferation assay,and plate clonal formation assay were used to detect the effect of AIM2 on the proliferation of H8 cells.Meanwhile,the effect of AIM2 on the migration ability of H8 cells was detected by Transwell assay.Results1.Immunohistochemical results showed that AIM2 was not expressed or weakly in normal tissues,but was highly expressed in CIN and squamous cell carcinoma compared with normal tissues,and the expression intensity was correlated with the degree of cervical lesions.RT-qPCR and Western blot results showed that AIM2 was highly expressed in H8,Hela,Siha and Caski cell lines(HPV+)compared with C33A cells(HPV-).2.ROC curve showed that AIM2 had good diagnostic value in LSIL and HSIL diagnosis.AIM2 and P16 have their own advantages and disadvantages in terms of diagnostic value.Combined diagnosis can maximize the diagnostic value.3.After the overexpression of AIM2,cck-8,Edu and plate clonal formation experiment results showed that the proliferation ability of H8 in cervical benign epithelial cells was enhanced and Transwell experiment results showed that the migration ability of H8 cells was enhanced.ConclusionAIM2 was highly expressed in cervical epithelial lesions and cervical cell lines in vitro with HPV infection,further verifying that HPV as cytoplasmic double-stranded DNA can activate AIM2.AIM2 has certain diagnostic value in distinguishing LSIL from HSIL,and its combination with P16 can maximize the diagnostic value.AIM2 can enhance the proliferation and migration of cervical benign epithelial cells.So AIM2 has the potential to promote malignant transformation of cervical benign epithelial cells.