MiR-1182 Inhibited Metastasis And Proliferation Of Ovarian Cancer By Targeting HTERT

Objective The current study found that microRNA RNA(miRNA)is closely related to the development of a variety of malignant tumors that are harmful to human health.It is involved in the regulation of biological characteristics of tumor cells,such as proliferation,invasion,metastasis,apoptosis,etc.The role of tumor suppressor genes or oncogenes suggests that miRNAs are expected to be targets for cancer therapy.MicroRNA 1182(miR-1182)is an important component of the microrna RNA regulatory network and plays an important regulatory role in many diseases.However,its development and mechanism of ovarian cancer has not been studied.The aim of this study was to investigate the expression of miR-1182 in the development,progression and metastasis of ovarian cancer,as well as its potential changes and targets,and provide a rational basis for clinical diagnosis and treatment of ovarian cancer.Methods In this study,we collected 50 specimens of ovarian epithelial cancer tissue from ovarian cancer patients between March 2015 and March 2018 in our hospital.A total of 50 specimens of normal ovarian tissue were collected.In the first part,the expression and changes of miR-1182 in ovarian cancer tissues,cells(SKOV3)and normal ovarian tissues and cells(IOSE80)were observed by real-time quantitative PCR(RT-PCR).In the second part,in order to study the potential targets of miR-1182,we used TargetScan,miRDB and microRNA three published algorithms to detect the possible targets of miR-1182 and use dual fluorescein.The enzyme reporter system clarified that human telomerase reverse transcriptase(hTERT)is a direct target of miR-1182,so hTERT caught our attention and was implemented in the next experiment.In the third part,we established three groups in SKOV3 cells for similar experiments(miR-NC group,miR-1182 mimic group and mimic + hTERT group),The effect of miR-1128 on the expression of target gene hTRET was detected by RT-PCR and Western blot,and the correlation between miR-1182 and its target gene was clarified.The effect of miR-1128 on the expression of the target gene hTRET,thereby clarifying the correlation between miR-1182 and its target gene.In the fourth part,we further tested the proliferation rate of ovarian cancer cells by MTT assay,and detected the migration and invasion ability of ovarian cancer cells by Transwell method.Results1.miR-1182 is down-regulated in ovarian cancer tissues and cells.We observed the expression of miR-1182 in ovarian cancer tissues and cells(SKOV3 cells)and normal ovarian tissues and cells(IOSE80 cells)by RT-PCR.The results showed that compared to the normal ovarian tissues,the expression level of miR-1182 in ovarian cancer tissues was statistically lower(p<0.0001,Figure 1-1).Compared with normal ovarian cells(IOSE80 cells),miR-in ovarian cancer cells(SKOV3 cells).The expression level of 1182 also showed a downward trend,and the difference was statistically significant(p<0.001,see Figure 1-2).Therefore,we believe that miR-1182 has a certain regulatory effect in the progression of ovarian cancer.2.hTERT is a direct target of miR-1182 in ovarian cancerUsing the publicly available algorithms of TargetScan,miRDB and microRNA,we hypothesized that human Telomerase Reverse Transcriptase(hTERT)is a potential target for miR-1182 and establishes a wild type containing hTERT 3'UTR and contains hTERT.The luciferase reporter vector of the 3'UTR mutant miR-1182 seed sequence uses mimics to increase the expression of miR-1182,resulting in a decrease in the luciferase activity of the wild-type hTERT 3'UTR reporter gene,but not for the mutant Effects(P<0.001,Fig.2-2),suggesting that hTERT expression can be affected by modulation of miR-1182.3?Correlation between miR-1182 and hTERTWe established three groups in SKOV3 cells for similar experiments(miR-NC group,miR-1182 mimic group and mimetic + hTERT group),and can be found by real-time PCR analysis and Western blot Western blotting,SKOV3 cells.Upregulation of miR-1182 reduced the expression level of hTERT(Fig.3-1,Fig.3-2),and the difference was statistically significant(P<0.05).These data indicate that hTERT may be negatively regulated by miR-1182.4.miR-1182 inhibits proliferation,invasion and migration of ovarian cancer cellsWe detected the cell proliferation rate by MTT assay.The MTT results showed that the cell proliferation rate of SKOV3 cells decreased after transfection of miR-1182 mimics.In contrast,down-regulation of miR-1182 promoted ovarian cancer cell growth(Fig.4-1).In Transwell experiments,we found that the migration and invasion ability of SKOV3 cells inhibited miR-1182 by up-regulating miR-1182,and the migration and invasion of ovarian cancer cells were enhanced by down-regulation of miR-1182(p<0.05,Fig.4-2,Fig.4-3)?Conclusions First,the abnormal expression of miR-1182 in ovarian cancer tissues and cells(SKOV3)was confirmed,which further confirmed that hTERT is a potential target of miR-1182,hTERT expression can be regulated by miR-1182,and hTERT is negative by miR-1182 regulation,which affects the proliferation and invasion and migration of ovarian cancer cells,reveals that the miR-1182/hTERT axis may be a potential target for the diagnosis and treatment of ovarian cancer.