| With the popularization of cytological screening and the expansion of cervical cancer vaccine application,cervical cancer prevention and early diagnosis have made some progress,but the metastasis and recurrence rate has not been controlled,and the mortality is still very high.Therefore,it is necessary to further clarify the pathogenesis of cervical cancer and provide new theoretical basis for the prevention and targeted intervention of cervical cancer.Micro RNA(miRNA)plays an important role in a variety of human malignant tumor diseases,including cervical cancer.Among them,miR-218 plays a role of tumor suppressor gene in the occurrence and development of cervical cancer,but its mechanism of inhibiting cervical cancer is not clear.Through database prediction,By database prediction,we suggest that transforming growth factor-β1(TGF-β1)may be the target gene of miR-218,These data suggest that miR-218 may be associated with TGF-β1in the regulatory mechanism of cervical cancer.Their association with cervical cancer has potential for targeted research.The purpose of this study was to elucidate the mechanism of miR-218 targeting TGF-β1 in regulating proliferation,apoptosis,migration and invasion of cervical cancer cells.Hela cells of cervical cancer were cultured and divided into up-regulated miR-218 group,down regulated miR-218 group,down regulated miR-218 and TGF-β1 inhibitor treatment group and control group.RT-PCR and Western blot were used to verify the expression and target regulation of TGF-β1 by miR-218,and the targeted binding sites of miR-218 and TGF-β1 were predicted by database and verified by dual-luciferase assay;then the changes of cell proliferation and migration were detected by CCK8,flow cytometry,scratch and Transwell;the mitochondrial membrane potential was detected by JC-1 staining,the ATP synthesis ability was determined by colorimetry,and the expression of mitochondrial apoptotic proteins such as Cytoc,Bcl-2,Caspase-3,Caspase-8 and Caspase-9 were detected by Western blot and luciferase.The results showed that miR-218 could bind to TGF-β1,and miR-218 could inhibit TGF-β1.In addition,miR-218 can inhibit the proliferation and invasion of Hela cervical cancer cells,and promote cell apoptosis;TGF-β1 inhibitor can reverse the promotion function of miR-218 silencing on proliferation and migration;moreover,miR-218 may regulate mitochondrial pathway through TGF-β1,increase the expression of Caspase-3,Caspase-9,Bax protein,reduce the expression of Bcl-2 protein,resulting in the decrease of mitochondrial membrane potential and ATP synthesis,promote the process of apoptosis.To further verify this conclusion,we used the nude mouse tumor model,set the same grouping treatment as the cells,and compared the survival rate,tumor volume and tumor size of mice.The results suggest that miR-218 can inhibit tumor formation in nude mice,and TGF-β1 inhibitor can reverse the promotion function of miR-218 silencing on tumor.miR-218 can regulate mitochondrial pathway through TGF-β1,increase the expression of Caspase-3,Caspase-9 and Bax protein,reduce the expression of Bcl-2 protein,reduce mitochondrial membrane potential and ATP synthesis ability,and promote the apoptosis process of tumor in vivo.In conclusion,miR-218 can inhibit the expression of TGF-β1 by targeting TGF-β1,so as to promote the mitochondrial apoptosis pathway and inhibit the proliferation and migration of cervical cancer cells in vitro and in vivo.This study revealed a new mechanism of miR-218 targeting TGF-β1 to regulate mitochondrial apoptosis and further clarified the regulatory mechanism of miR-218 in the occurrence and development of cervical cancer,so as to provide a reference for clinical diagnosis and targeting of cervical cancer intervention,besides,provides a new theoretical basis and a new target for the recurrence,metastasis and precise treatment of cervical cancer,which has important scientific significance and clinical application value in the early diagnosis,early intervention,treatment and prognosis evaluation of cervical cancer. |
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