| Objective:Cervical cancer is a common cancer in women and the fourth leading cause of cancer-related death,posing a serious threat to women's health.IRhoms is a non-catalytic protein of Rhomboid protease family.IRhoms has been confirmed as a carcinogenic factor for a variety of tumors,but it is not clear whether iRhoms is related to cervical cancer(CC)and its mechanism of action.The purpose of this study is to clarify the relationship between the expression of iRhoms and the clinicopathological classification and prognosis of cervical cancer,observe the effect of iRhoms on the proliferation,cell cycle distribution and apoptosis of Hela cells in cervical cancer,and to explore the mechanism of Wnt pathway in cervical cancer.Methods:1.Twenty fresh cervical cancer tissue samples and paired adjacent non-cancerous cervical tissue samples were collected.The expression of iRhoms was detected by western blot,and the expression differences in normal cervical and cervical cancer were compared.Eighty-three patients with cervical cancer at the Ia2-IIa2 stage were collected,and the relationship between the expression of iRhoms protein and the clinicopathological type was detected by immunohistochemical method.Kaplan-meier and Cox regression analysis were performed to analyze the relationship between iRhom1,iRhom2 and ki-67 and the survival rate of cervical cancer patients.2.Cervical cancer Hela cell lines were cultured,transfected with iRhoms sh RNA-encoded lentivirus,iRhoms expression was knocked down,and functional experiments were conducted at the cell level by MTT assay,clonal formation assay,flow apoptosis assay and cell cycle assay.3.Will knock out iRhom2 cervical cancer cells and the negative control group by genome microarray detection,and use the Ingenuity Pathway Analysis(IPA)system(Ingenuity Systems,Mountain View,CA,USA)for data Analysis,screening iRhoms possible pathways in cervical cancer and related genes,using western blot detection related protein expression profile of functional verification.Results:1.Compared with adjacent normal tissues,the expression levels of iRhom1and iRhom2 in cervical cancer tissues were higher,and the strong expression levels of iRhom1,iRhom2 and Ki67 were significantly correlated with staging,tumor size and para-uterine infiltration.2.Transfection with iRhoms sh RNA-encoded lentivirus was shown to inhibit proliferation,promote apoptosis and interfere with cell cycle.3.Genomic microarray detection and IPA data analysis showed that iRhom2 knockout changed many tumor-related signal transduction pathways and oncogenes,and Wnt / ?-catenin pathways were screened out.Western blot results showed that iRhom1 knockdown significantly inhibited the expression of ?-catenin,Myc,TGFBR2 in Hela cells,and increased the expression of FAS.Knockdown of iRhom2 significantly inhibited the expression of ?-catenin,GSK3 and Myc in Hela cells,which was consistent with the genomic microarray data.Conclusion:The expression of iRhoms is increased in cervical cancer,and the expression is related to clinical characteristics and prognosis.Patients with high expression of iRhom1 and iRhom2 have a short progression-free survival period and a low total survival period.Knockdown of iRhom1 and iRhom2 can promote apoptosis of cervical cancer cells,inhibit cell proliferation,interfere with cell cycle,and verify multiple genes and cancer-related pathways involved in this process.The results of this study indicated that iRhoms,as an oncogene,may be a potential tumor marker related to diagnosis and prognosis,and may become a new therapeutic target for cervical cancer. |
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