Study On The Role Of EDIL3 Gene In The Development Of Cervical Cancer

Objective: To explore the expression levels of EDIL3 in tissues and cells of cervical cancer and the correlation between the expression of EDIL3 and HPV infection.To explore the effects of EDIL3 on the cell proliferation,migration,cell cycle and epithelial–mesenchymal transition(EMT)of cervical cancer cells Si Ha and He La,and analyze the molecular mechanism of EDIL3 affecting the proliferation and migration of cervical cancer cells Si Ha and He La.Methods:(1)Bioinformatics databases GEO,GEPIA,UALCAN,and Onco Lnc were used to analyze the expression level of EDIL3 gene in cervical cancer tissue,the methylation status of gene promoter,and prognostic significance.(2)The expression levels of EDIL3 protein and m RNA in tissues and cells of cervical cancer were detected by immunohistochemistry,Western blot and q RT-PCR.(3)Whether the cervical cancer tissue is infected with HPV is detected using the second generation hybridization capture technique(HC-2).(4)EDIL3 overexpression plasmid and RNA interference plasmid were designed and divided into experimental group(overexpression of EDIL3 plasmid or RNA interference knockdown plasmid)and control group NC(empty plasmid).The proliferation of cervical cancer cells Si Ha and He La were assessed by using MTT test and colony formation assay,and the migration of cervical cancer cells Si Ha and He La were assessed by using transwell test and cell scratch test.The expression levels of EMT-related molecules in cervical cancer cells Si Ha and He La were determined by using Western blot.(5)Flow cytometry is used to analyze the cell cycle changes of cervical cancer cells after the EDIL3 is overexpressed.(6)The concentration of EDIL3 protein in the culture supernatant of cervical cancer cells after overexpression of EDIL3 gene was detected by ELISA.(7)The FAK-Erk pathway related molecular protein expression level were determined by Western blot.Results: Bioinformatics database analysis showed that EDIL3 m RNA was low expression and promoter hypermethylation in cervical cancer tissues(P < 0.05),and there was no difference in expression levels among different stages of cervical cancer(P > 0.05),High EDIL3 expression was associated with poor prognosis of cervical cancer patients(P < 0.05).Immunohistochemical staining showed that the expression level of EDIL3 protein in the cytoplasm of cervical squamous cell carcinoma tissue cells was lower than that of normal cervical tissue(P < 0.05),and the expression level of EDIL3 protein was significantly correlated with HPV infection(P < 0.05).Compared with normal cervical tissue,the expression level of EDIL3 protein increased in the intercellular matrix of cervical squamous cell carcinoma(P < 0.05),but there was no significant correlation between the expression level of EDIL3 protein and HPV infection(P > 0.05).The EDIL3 protein and EDIL3 gene m RNA expression level of cervical cancer Si Ha and He La cells line compared with the Ha Ca T cells line was significantly decreased by Western blot and q RT-PCR,the difference is statistic significance(P < 0.05).The cervical cancer cells Si Ha and He La were successfully transfected with the overexpressed EDIL3 plasmid and the RNA interfering plasmid.The results of the MTT experiment and colony formation assay showed that compared with the control group NC,the proliferation of cervical cancer cells in the overexpressed EDIL3 experimental group was significantly increased(P < 0.05),while the proliferation of cervical cancer cells in the RNA interference EDIL3 experimental group was significantly decreased(P < 0.05).The results of cell scratch test and transwell test showed that compared with the control group NC,the migration ability of cervical cancer cells in the overexpressed EDIL3 experimental group was significantly increased(P < 0.05).At the same time,Western blot results showed that the expression level of EMT molecule E-cadherin in the overexpressed EDIL3 experimental group was decreased,and the expression levels of vimentin,Slug andβ-catenin were increased(P < 0.05).Cell cycle results showed that after the overexpression of EDIL3,the cell cycle time of cervical cancer cells Si Ha and He La was shortened,and the cell proliferation was increased(P < 0.05).ELISA results showed that compared with the control group NC,the experimental group overexpressing EDIL3 in cervical cancer cells Si Ha and He La had significantly increased EDIL3 protein concentration in the cell culture supernatant(P < 0.05).Western blot results showed that After the overexpression of EDIL3,the molecular protein expression level of FAK-Erk signaling pathway was significantly increased.(P < 0.05).Conclusion: The expression level of EDIL3 protein in the cytoplasm of cervical squamous cell carcinoma histiocyte was lower than that of normal cervical histiocyte,and the expression level of EDIL3 protein was correlated with HPV infection;The expression level of EDIL3 protein in the stroma of cervical squamous cell carcinoma histiocyte was higher than that of normal cervical histiocyte.Overexpression of EDIL3 gene can promote the proliferation and migration of cervical cancer cells,and RNA interference knockdown of EDIL3 gene can inhibit the proliferation of cervical cancer cells.Overexpression of EDIL3 gene may affect the proliferation and migration of cervical cancer cells through up-regulation of FAK-Erk signaling pathway.