Investigate The Relationship Between PBX3 Expression Level And Clinicopathological Factors Of Cervical Cancer, And Further Study On Molecular Biological Mechanism

Objective: Cervical cancer(CC)is the second most prevalent gynecologic malignancy in the world.Although the methods of diagnosis and therapy of CC have been developed in recent years,the overall prognosis is still not satisfactory.How to develop new treatment methods including gene therapy is one of the key problems to improve the curative effect of cervical cancer.The Pre-b-cell leukemia homeobox 3(PBX3)was a gene raised expression in many malignancies,but its role in cervical cancer was not clear.In this study,the expression level,clinical significance,molecular biological function and mechanism of PBX3 in cervical cancer were discussed to clarify its potential as a new diagnostic and therapeutic target for cervical cancer.Methods: By real-time polymerase chain reaction(RT-PCR),Western blotting and immunohistochemical staining were used to detect the expression level of PBX3 in a variety of cervical cancer cell lines and clinical specimens,and then to analyze the clinicopathological features and survival rates associated with PBX3 expression levels.RNA interference(siRNA)was used to inhibit the expression level of PBX3 and to identify the inhibitory effect of PBX3 expression in each cervical cancer cell line.The effect of PBX3 on the proliferation of cervical cancer cells was investigated by MTT assay,soft agar clone formation assay and plate cloning assay.Then,we established a model of cervical cancer subcutaneous transplantation model with each cervical cancer cell line to explore the relationship between PBX3 inhibition and the growth curve and tumor weight of cervical cancer models.Finally,the signaling pathway and molecular biological mechanism of PBX3 regulating cervical cancer proliferation dependence were discussed.Results: The study confirmed that PBX3 mRNA and protein expression level were significantly increased in nearly all of the cervical cancer cell lines compared with normal cervical cell line.It was found that the expression level of mRNA and protein level of PBX3 in cervical cancer tissue was significantly higher than that of paired normal cervical tissue.After analyzing the relationship between the expression level of PBX3 expression level with clinicopathological features,it was found that the expression level of PBX3 was correlated with tumor diameter,pathological grading,lymph node metastasis,infiltration depth,vascular infiltration and clinical staging.Multivariate analysis showed that PBX3 was one of the independent predictors of poor prognosis in patients with cervical cancer.Survival analysis showed that the overall survival rate of patients with high expression PBX3 was lower compared with those with low expression PBX3.In vitro experiments showed that the 2 kinds of PBX3-siRNA could reduce the expression level of PBX3 protein in various cervical cancer cell lines(Hela and Siha),and the reduction of PBX3 expression could inhibit the proliferation ability and cloning ability of cervical cancer cell lines(Hela and Siha).In vivo studies also found that in PBX3 reduced cell lines,in vivo tumor growth and tumor weight were significantly inhibited.The inhibition effect was produced by PBX3 protein expression level.Finally,the mechanism of PBX3 in cervical cancer cells was discussed,and it was found that the expression level of p-AKT protein decreased stably after decreasing PBX3 expression level,and PBX3 could produce positive regulation effect on cervical cancer cell proliferation by activating AKT pathway.Conclusion: PBX3 regulating proliferation of cervical cancer through the AKT pathway.PBX3 may be the prognostic marker of cervical cancer,and is expected to become a new target for cervical cancer gene therapy.