TCONS₀0026907,Acts As CeRNA,Is Involved In The Development Process Of Cervical Cancer

BackgroundCervical cancer is one of the most common gynecologic cancers and occupies a large proportion in cancer deaths among women worldwide.Long non-coding RNAs?lncRNAs?,a kind of non-coding RNA longer than 200 nucleotides,are closely related to tumor genesis.MicroRNAs?miRNAs?,a kind of non-coding RNAs with about 20 nucleotides,play important roles in various diseases including human cancers.ELK 1 is a member of the Ets family of transcription factors and of the ternary complex factor?TCF?subfamily.We screened cervical cancer related lncRNAs using microarray experiments,then qRT-PCR further proved the expression levels of lncRNAs,and speculated that lncRNA TCONS₀0026907 which may serve as ceRNA,is involved in the progression and prognosis of cervical cancer through inhibition of miR-143-5p and promotion of ELK1.LncRNA TCONS₀0026907 may be potential biomarkers for the diagnosis of cervical cancer.Objectives:To screen cervical cancer related lncRNAs and to explore the potential role of lncRNA TCONS₀0026907 in the process of development cervical cancer through miR-143-5p and ELK1.Methods:Firstly,we have screened cervical cancer related lncRNAs using microarray experiments,the mRNA expression level of TCONS₀0026907 was detected by qRT-PCR in cervical cancer tissues,and the survival rate of patients was analyzed.Secondly,The proliferation,cycle distribution,apoptosis,migration and invasion abilities of cervical cancer cells were detected by CCK-8 assay,flow cytometry,migration and invasion assay respectively.Thirdly,qRT-PCR assay detected the mRNA expression level of ELK1 and miR-143-5p.The protein expression levels of ELK1,p-ELK1,C-fos,Cyclin D1,and Bcl-2 were measured by Western blotting.We constructed the wild type TCONS₀0026907 and mutant TCONS₀0026907 luciferase reporter,and the wild type ELK1 or mutant ELK1 luciferase reporter respectively,and were co-transfected into HeLa cells with miR-143-5p.The luciferase activity was measured using the luciferase report gene assay.p-ELK1 expression was detected by the immunofluorescence assay.Observed the change of cervical cell cycle,proliferation,migration,invasion and apoptosis when overexpress miR-143-5p or silence ELK1.Furthermore,athymic mice were injected the transfected Hela and SiHa cells to making cervical cancer models.The growth curve and the tumor volume were detected in tumor formation assay.The expression levels of ELK1,p-ELK1,C-fos,Cyclin D1,and Bcl-2 were measured in vivo.ResultsMicroarray experiments had screened 5 cervical cancer related lncRNAs.The results showed that TCONS₀0026907 expression level was higher in cervical cancer tissues than adjacent noncancerous tissues.The survival rate was lower in high expression group than low expression group.The high expression group included a significantly larger tumor size,a higher TNM tumor stage and a stronger possibility of cancer metastatic when compared with the low expression group.In addition,we found that TCONS₀0026907 expression level was higher in HeLa cells and SiHa cells than in CaSki and C33A cells.Silence of TCONS₀0026907 expressions by lentivirus inhibited cervical cell cycle,proliferation,migration and invasion,and promoted apoptosis.Silence of TCONS₀0026907 expression suppressed the growth of cervical tumor and regulates ELK1,p-ELK1,C-fos,Cyclin D1,and Bcl-2 expression in vivo and in vitro.We also found TCONS₀0026907 negatively regulated miR-143-5p and miR-143-5p negatively regulated ELK1.Furthermore,overexpressed miR-143-5p or silence ELK1 can inhibite cervical cell cycle,proliferation,migration and invasion,and promote apoptosis.Conclusions1.TCONS₀0026907 mRNA expression was significantly higher in cervical cancer tissues and the survival rate was lower in high expression group than low expression group.2.Silence of TCONS₀0026907 by lentivirus inhibited cervical cell cycle,proliferation,migration and invasion,and promoted apoptosis.Silence of TCONS₀0026907 expression suppresses the growth of cervical tumor and regulates ELK1,p-ELK1,C-fos,Cyclin D1,and Bcl-2 expression in vivo.3.TCONS₀0026907 served as ceRNA inhibited miR-143-5p expression and accelerates ELK1 expression and participated in the process of tumor development.4.TCONS₀0026907 may become a very important class of biomarker in patients with cervical cancer.Novelty1.This research firstly reported that lncRNA TCONS₀0026907 was closely associated with cervical cancer,and deeply discussed its role and mechanism in development process of cervical cancer.2.LncRNA TCONS₀0026907 maybe become a new tumor markers of the cervical cancer.