| Objective: Cervical cancer is one of the most common cancers in women and the fourth leading cause of cancer deaths in women worldwide.The persistent infection of HR-HPV(High risk human papilloma virus)is closely related to the occurrence of cervical cancer,which may lead to a series of biological changes such as SNP variation,fragment deletion or duplication,activation of proto-oncogenes and inactivation of tumor suppressor genes in host genes.At present,the molecular mechanism of the occurrence,development and metastasis of cervical cancer is still unclear.The study of the genes and functions related to cervical cancer is conducive to the exploration of the specific molecular mechanism of cervical cancer,the development of biomarkers and the development of new targets for the diagnosis and treatment of cervical cancer.PABPC1(poly real binding protein cytoplasmic 1)is A member of the family of PABP RNA binding protein,can regulate RNA stability related protein complexes,and m RNA metabolism,cell growth and development of tumor,and many other important life activities.PABPC1 is expressed in a variety of tumor tissues and is closely related to tumor growth and metastasis.However,the molecular mechanism of its action remains unclear.In this study,transcriptome study found that PABPC1 was up-regulated in cervical squamous cell carcinoma,and it was speculated that PABPC1 might be involved in the regulation of the occurrence and development of squamous cell carcinoma.This study intends to discusses PABPC1 expression in cervical squamous carcinoma tissues,analysis the relationship between its expression and clinical pathological parameters related,and through the SiRNA interference technology PABPC1 silence gene expression,the lower expression was observed after cervical squamous cell carcinoma of the SiHa cell proliferation,migration and other biological behavior change,explores PABPC1 expression change affects the mTOR and AKT/MAPK signaling pathways regulate the molecular mechanism of tumor cells,and to clarify the occurrence of cervical cancer provide new theoretical basis.Methods: 1.To investigate the expression of PABPC1 in cervical squamous cell carcinoma tissues and its clinical significance:Immunohistochemistry was used to detect the distribution and expression of PABPC1 protein in 42 cervical squamous cell carcinoma tissues and para-cancerous tissues,and the correlation between the expression of PABPC1 in cancer tissues and various clinicopathological features was analyzed.2.To explore the effect of silencing PABPC1 gene on the biological behavior of SiHa cells in vitro:(1)SiHa cells were transfected with SiRNA interference technology,and the expression changes of PABPC1 m RNA and protein were detected by Real-time PCR(RT-PCR)and Western blot assay to verify the transfection efficiency.(2)CCK8 and plate cloning formation experiments were used to compare the changes of proliferation and cloning formation ability of SiHa cells before and after PABPC1 gene silencing.(3)Scratch test and Transwell test were used to detect the effect of silencing PABPC1 gene on the migration and invasion ability of SiHa cells in vitro.3.To investigate the molecular mechanism of silencing PABPC1 affecting mTOR and Akt/MAPK signaling pathway in SiHa cells: Western blot assay was used to detect the effects of silencing PABPC1 gene on the expression of key proteins including AKT1,RPS6,ERK1/2,RKS1P90 and Rab11 in mTOR and AKT/MAPK signaling pathway in SiHa cells.Real-time PCR(RT-PCR)assay was used to detect the m RNA expression of mTOR and ERK.Results: 1.Expression and clinical significance of PABPC1 in cervical squamous cell carcinoma tissues: Immunohistochemical results of 42 cervical squamous cell carcinoma samples showed that the expression of PABPC1 in cervical squamous cell carcinoma tissues was significantly higher than that in adjacent tissues(P<0.01);The positive expression of PABPC1 was statistically correlated with FIGO stage of cervical squamous cell carcinoma(P<0.05),but not with age,tumor size,degree of tissue differentiation and lymph node metastasis(P>0.05).2.Effects of silencing PABPC1 gene on proliferation,clone formation,migration and invasion ability of SiHa cells in vitro:(1)SiHa cell line system with instantaneous silencing of PABPC1 was established,and the validation results showed that the silencing efficiency of SiRNA-1 and SiRNA-2 fragments of PABPC1 on SiHa cells met the experimental requirements.(2)CCK8 and plate cloning formation experiments showed that the proliferation and cloning formation ability of SiHa cells were significantly inhibited and decreased after PABPC1 silencing compared with the control group.(3)Cell scratch and Transwell assay results showed that the in vitro migration and invasion ability of SiHa cells were significantly reduced after PABPC1 gene silencing compared with the control group.3.The molecular mechanism of regulating mTOR and AKT/MAPK signaling pathways after the silencing of PABPC1 in SiHa cells: Real-time PCR(RT-PCR)and Western blot results showed that the expression levels of mTOR m RNA and its phosphorylated protein were significantly decreased after the silencing of PABPC1.The level of ERK m RNA in Akt/MAPK signaling pathway and the expression of ERK1/2phosphorylated Y204/187 protein were down-regulated.Conclusion: 1.The expression of PABPC1 is up-regulated in cervical squamous cell carcinoma tissues,and its high expression is correlated with FIGO staging of cervical carcinoma.2.Silencing of PABPC1 gene can inhibit the proliferation,cloning formation,migration and invasion of SiHa cell line in vitro,so PABPC1 may be a cancer driver gene.3.PABPC1 functional protein may regulate mTOR expression through activation of ERK in MAPK signaling pathway,thus promoting the occurrence of cervical squamous cell carcinoma. |
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