The Research Of CENPU Regulating The Proliferation,Migration And Invasion Of Cervical Cancer Cells Through PI3K/AKT Signaling Pathway

Cervical cancer(CC),as one of the most common malignant tumors among women worldwide,recurrence and metastasis are important causes of death,The occurrence and continuous progression of precancerous lesions in cervical eventually lead to the occurrence of cervical cancer.Due to the unclear understanding of the developmental pattern of normal cervical tissues progression to carcinogenesis.the study of the molecular mechanisms of this condition will provide a scientific basis for the prevention and treatment of cervical cancer.It has been confirmed that the stability of Centromere protein U(CENPU)is a key factor in cell proliferation and division,It has been reported that CENPU is overexpressed in breast cancer,ovarian cancer and other tumors,and in vitro cell experiments have confirmed that its abnormal expression can participate in the proliferation,migration and invasion of tumor cells through phosphatidylinositol 3-kinase(PI3K)/ protein kinase B(AKT)pathway,but it is rarely reported in cervical cancer.In view of this,this study used immunohistochemistry(IHC)staining to explore the expression significance of CENPU in cervical intraepithelial lesions and cancerous tissues and its relationship with clinical pathological characteristics.The effects on proliferation,migration and invasion of cervical cancer cells were studied by cell experiment.47 cases of normal cervical paraffin block,21 cases of LSIL paraffin block,30 cases of HSIL paraffin block and 64 cases of CC paraffin block treated in the First Affiliated Hospital of Hebei North University from January 2019 to December 2021 were selected.The expression of CENPU was detected by IHC,and Analyzing the correlation between its expression and clinical pathological data of cervical cancer.si RNA transfection technique was used to explore the influence of silenced CENPU expression on the biological behavior of cervical cancer Si Ha and He La cells in vitro: Cells were divided into MOCK group,NC group and experimental group.Real-time fluorescent quantitative polymerase chain reaction(RT-q PCR)and Western Blot(WB)were used to validate the silence efficiency.The cell proliferation ability of CENPU silenced cells in vitro was detected by CCK8;The migration ability of CENPU silenced cells in vitro was detected by scratch assay;the cell clone formation ability of CENPU silenced cells in vitro was detected by Plate clone formation assay;The invasion ability of CENPU silenced cells in vitro was detected by Transwell assay.To explore the possible mechanism of the change of cervical cancer cell function after changing CENPU expression,PI3 K,AKT,p-PI3 K,PAKT protein expression of PI3K/AKT signaling pathway and EMT related proteins such as N-cadherin,Vimentin,E-cadherin were detected by WB.The results of our study showed that: CENPU was expressed in normal cervical tissues,CIN tissues and cervical cancer tissues,and the expression level increased with the progression of the lesions.The expression intensity of CENPU in cervical intraepithelial neoplasia(CIN)and cervical cancer tissues was higher than that in normal cervical tissues(P<0.05),the expression intensity of HSIL group was significantly higher than that of LSIL group(P <0.05),but there was no significant difference between cervical cancer group and HSIL group(P>0.05),The relative expression of CENPU in cervical cancer tissues was not correlated with age,histological type,differentiation degree,clinicopathologic stage,lymph node metastasis and invasion depth(P>0.05).q RT-PCR and WB experiments confirmed that the relative expression levels of CENPU m RNA and protein in the experimental group were significantly lower than those in the control group after CENPU expression silencing(P<0.05).Cell function experiment results showed that the proliferation ability,scratch healing rate,cell clone formation ability and invasion ability of cervical Si Ha and He La cells were decreased after CENPU expression was silenced.Meanwhile,WB results showed that in the CENPU silencing experimental group,the expression of p-PI3K/PI3 K,p-AKT/AKT,EMT-related proteins N-cadherin and Vimentin was significantly lower than that of the control group,while the expression of E-cadherin was significantly higher than that of the control group(P<0.05).It is concluded that the abnormal high expression of CENPU protein promotes the progression of cervical intraepithelial lesions and cervical cancer;CENPU can affect the proliferation,migration and invasion of cervical cancer cells through PI3K/AKT signaling pathway,and induce tumor cells to develop EMT to regulate the malignant biological behavior of cervical cancer cells.