Expression Of IFRD1 And Its Significance In Different Cervical Lesions

Objective:By Observing the expression of IFRD1 protein and its m RNA in different cervical lesions and in the tissues of cervical squamous cell carcinoma(SCC)with different clinical and pathological parameters,this study aims to research the the relationship between IFRD1 gene and the pathogenetic and progression mechanisms of cervical lesions,to explore new biomarkers and gene therapy targets for the prevention,diagnosis and treatment of cervical lesions.Methods:There were 116 cervical lesion tissues collected from different clinical cases,which included 20 cases of chronic cervicitis(control group),20 cases of low-grade squamous intraepithelial lesion(LSIL group),20 cases of high-grade squamous intraepithelial lesion(HSIL group),48 cases of cervical squamous cell carcinoma(SCC group),8 cases of cervical adenocarcinoma(AUC group).The expression of IFRD1 protein in paraffin sections of different cervical lesion tissues and its m RNA in fresh tissues were detected respectively by Immunohistochemistry(IHC)and real-time quantitative PCR(Qrt-PCR).Using the two methods mentioned above,this study aims to firstly compare the differences in expression of IFRD1 protein and its m RNA among chronic cervicitis tissue,LSIL tissue,HSIL tissue and SCC tissue.Secondly,the aim is to research the differences of IFRD1protein and its m RNA among the tissues with different clinical and pathological parameters.Thirdly,this study aims to analyze the expression of IFRD1 m RNA differences between AUC tissues and chronic cervicitis tissues and the expression of IFRD1 protein in AUC tissues and normal cervical gland tissues.The expression of IFRD1protein in chronic cervicitis tissues collected from 6 clinical cases and SCC tissues from other 6 clinical cases were detected by Western blotting(WB)for further verification.According to the level of cervical lesions,clinical pathological parameters,the experimental data were analyzed in different aspects.The statistical results were analyzed by statistical software SPSS 20.0.The difference between groups was statistically significant(P<0.05).Results:1.It was detected by IHC that IFRD1 protein expressed mainly in the squamous epithelium in chronic cervicitis tissue,LSIL tissue and HSIL tissue,which were brown granules located in the cytoplasm,and the color of each group had no significant difference.In Chronic cervicitis tissues,IFRD1 protein expressed mainly in the basal layer and sub-basal layer of squamous epithelium.In the CIN tissues,IFRD1 protein expressed to varying degrees in the squamous epithelium,and the expression level thickened with the severity of cervical lesions.In the SCC cancer nest tissues,IFRD1 protein diffusely expressed in all layers,which showed a small amount of dotted expression in the tissue outside cancer nest and the interstitial outside the cervical squamous epithelium.The results of the rank sum test and Spearman correlation analysis showed that the expression of IFRD1 protein in control group,LSIL group,HSIL group and SCC group showed a trend of increasing(χ~2=83.44,P=0.000,P<0.05,r=0.884).Compared between groups,the expression of IFRD1 protein gradually increased with the severity of cervical lesions,SCC group is stronger than HSIL group,HSIL group is stronger than LSIL group,and LSIL group is stronger than Control group.The difference between the groups was statistically significant(P<0.05).In AUC and glandular cytoplasm in normal cervical glandular tissue,IFRD1 protein was expressed in varying degrees.The glandular cytoplasm of AUC tissues expressed more and was dark brown.Using t-test for statistical analysis,the expressio diffuse of IFRD1 protein in AUC tissues was higher than that in normal cervical tissues,and the difference was statistically significant(P=0.030,P<0.05).2.It was detected by Qrt-PCR that the relative expression of IFRD1 m RNA was 1.40±0.39 in the Control group,2.93±0.94 in the LSIL group,5.37±1.56 in the HSIL group,11.32±2.59 in the SCC group and 11.04±2.62 in the AUC group.One-way analysis of variance showed significant differences between groups(F=131.78,P=0.000,P<0.05).Using dunnett-t test for statistical analysis between each group,the expression levels of IFRD1 m RNA in LSIL group,HSIL group,SCC group and AUC group were higher than those in control group,and the expression in LSIL group,HSIL group,SCC group and AUC group showed a gradual increase trend with the severity of lesion.Comparing between the groups,P values were<0.05,the difference was statistically significant.3.It was detected by WB that the relative expression of IFRD1 protein in SCC tissues was higher than that in chronic cervicitis tissues,and the difference was statistically significant by T-test(P=0.0002,P<0.05).4.In the IHC and Qrt-PCR detection results in the SCC tissues,Univariate analysis of variance(ANOVA)was used to analyze the data of different pathological grades and clinical stage.The t-test was used to analyze the data of myometrial invasion,vascular invasion,nerve invasion and pelvic lymph node metastasis.Compared among the SCC groups with different clinical and pathological parameters,the difference of IFRD1 and IFRD1 m RNA expression levels have no statistically significance(P>0.05).Conclusion:1.The expression of IFRD1 protein and the relative expression of IFRD1 m RNA increased with the severity of cervical lesions,which suggests that IFRD1 gene may play an important role in the pathogenesis of CIN and SCC.2.In IHC detection,with the lesion increasing,IFRD1 protein thickened from the basal layer of the cervical squamous epithelium to the surface gradually.In the SCC cancer nest tissues,IFRD1 protein diffusely expressed in all layers,suggesting that the gene may be involved in the proliferation of cervical squamous epithelial cells and SCC cells.3.In the SCC tissues,the expression level of IFRD1 protein and m RNA was not significantly correlated with the clinical pathology parameters of SCC(clinical stage,degree of tumor differentiation and pathological invasion).The present study suggests that IFRD1 gene is probably not involved in the invasion,metastasis,progression and prognosis of SCC.4.Both in the normal cervix glands and the cytoplasm of the AUC gland,IFRD1protein was expressed,and the expression intensity in AUC was higher than that in normal cervical gland group.Similarly,the expression intensity of IFRD1 m RNA in AUC group was higher than that in Control group,The present research reveals that IFRD1 gene may play a certain role in the pathogenesis of AUC.