MiR-424-5p Regulates The Proliferation,migration And Apoptosis Of Cervical Cancer Through The GPR30/PI3K/AKT/mTOR Signaling Pathway

Cervical cancer is one of clinical common tumors in women,pose a serious threat to the health of patients among them,the problem such as recurrence,metastasis and spread is a major cause of death in patients.Currently,cervical cancer treatment is surgery and radiation therapy,although the vast majority of patients with early cervical cancer can achieve the effect of cure,but due to the occurrence and progress of molecular mechanism is unclear,with advanced or metastatic cervical cancer,there is no satisfactory treatment.Therefore,it is urgent to find some better targeted therapeutic strategies to save more lives.Micro ribonucleic acid(micro RNA,mi RNA)plays an important role in regulating cell proliferation,differentiation and apoptosis,and it is closely related to the formation of malignant tumors.Many mi RNAs have been found to be abnormally expressed in cervical cancer,and are involved in the development of cervical cancer by targeting and regulating the downstream target proteins,and can also be used as potential biomarkers for the diagnosis of cervical cancer.Studies have shown that mi R-424-5p is involved in the proliferation,invasion and migration of a variety of tumors.However,there are few reports on the role and mechanism of mi R-424-5p in the occurrence and development of cervical cancer.G protein-coupled receptor 30(GPR30)is a novel estrogen receptor.On the one hand,GPR30 can specifically bind to estrogen and promote tumor progression by activating mitogen-activated protein kinase(MAPK)or phosphatidylinositol 3-kinase(PI3K)signaling pathways.On the other hand,GPR30 accelerates angiogenesis by regulating epidermal growth factor receptor(EGFR),and promotes the invasion and migration of tumor.Our previous study showed that GPR30 was highly expressed in cervical cancer.Is mi R-424-5p involved in the pathogenesis of the malignant biological behavior in cervical cancer? Does mi R-424-5p promote the occurrence and development of cervical cancer by targeting GPR30? What is the specific mechanism? It needs to be verified by further study.In this study,we investigated the effect of mi R-424-5p on the malignant biological behavior of cervical cancer He La cells by overexpressing or inhibiting the expression of mi R-424-5p.To observe the targeting relationship between mi R-424-5p and GPR30,and explore its mechanism preliminarily.The aim is to further search for new target genes and regulatory mechanisms,and provide a potential theoretical basis for targeted therapy of cervical cancer.Cervical cancer He La cells were divided into NC mimics group(transfected with mi R-424-5p mimics negative control),mimics group(transfected with mi R-424-5p mimics),NC inhibitor group(transfected with mi R-424-5p inhibitor negative control),inhibitor group(transfected with mi R-424-5p inhibitor).Real-time quantitative polymerase chain reaction(q RT-PCR)was used to assess the transfection effect.Changes of the morphology of He La cells after transfection under inverted microscope and scanning electron microscope(SEM).The proliferation,clonality,invasion,migration,cell cycle and apoptosis of cervical cancer cells were detected by Cell Counting Kit-8(CCK-8),plate colony formation experiment,Transwell invasion assay,wound healing assay and flow cytometry,respectively.The target gene of mi R-424-5p was predicted by the target gene prediction software,and verified by double luciferase reporter gene assay.The targeting relationship between mi R-424-5p and GPR30 were further assessed by q RT-PCR and Western blot(WB).The protein expressions related to PI3K/protein kinase B(AKT)/mechanistic target of rapamycin(m TOR)signaling pathway were assessed by WB.It was found that overexpression of mi R-424-5p decreased the number of He La cells,as well as the microvilli and pseudopod on the surface of the cells.Besides,the proliferation,clonality,invasion and migration ability of He La cells in the mimics group were significantly lower than that in the NC mimics group,while the inhibitor group were significantly higher than that in the NC inhibitor group.The proportion of G0/G1 in the mimics group was significantly higher than that in the NC mimics group,and the proportion of S and G2/M in the mimics group were significantly lower than that in the NC mimics group,while the inhibitor group was promoted the opposite results.The apoptosis rate in the mimics group was significantly higher than that in the NC mimics group,while the inhibitor group were significantly lower than that in the NC inhibitor group.we preliminarily verified GPR30 as a direct target gene of mi R-424-5p by target gene prediction and double luciferase experiment.The expression level of mi R-424-5p in cervical cancer tissues was significantly lower than that in normal cervical tissues,while GPR30 was higher than that in the NC mimics group.The expression of GPR30,phosphorylated protein kinase B(p-AKT)/AKT and phosphorylated mechanistic target of rapamycin(p-m TOR)/m TOR in the mimics group were significantly lower than that in the NC mimics group,while the inhibitor group were significantly higher than that in the NC inhibitor group,the difference of the results were significantly(all P<0.05 or P<0.01)In conclusion,the low expression of mi R-424-5p may promote the occurrence and development of cervical cancer,while the overexpression of mi R-424-5p can significantly inhibit the proliferation,clonality,invasion,migration,cell cycle of cervical cancer cells,and induce cell apoptosis.In summary,mi R-424-5p may affect the malignant biological behavior of He La cells by regulating the expression of GPR30 through PI3K/AKT/m TOR signaling pathway,and mi R-424-5p may become a potential target for cervical cancer gene therapy.