The Effects Of MiRNAs In Testes Specific Protease 50 High-expressing Tumor Cells

The testes-specific protease 50(TSP50)can paly a very important role in promoting tumorigenesis as a novel threonine protease.Except for its expression in normal testes,TSP50 is abnormally reactivated in many malignant breast tumor samples,in addition,knockdown of TSP50 inhibits cell proliferation and induces apoptosis in p19 cells.These results make TSP50 an attractive molecular marker and a potential target for diagnosis and therapy.Therefore,to clarify the regulatory mechanism of TSP50 expression is very important for cancer therapy.miRNAs are endogenous non-coding RNAs approximately 22 nucleotides in length that play an important role in the negative regulation of gene expression at the post-transcriptional level.Experimental evidence indicates that miRNAs are aberrantly expressed in different tumour types and implicated in multiple malignancy-related processes including cell proliferation,apoptosis,invasion and metastasis.In present study,we screened miRNAs that targeting TSP50 mRNA 3'UTR and examined their function on the cell proliferation,mobility and invation of TSP50 high-expressing cells.First we predicted four mi RNAs targeting TSP50 mRNA 3'UTR by using bioinformatics software and TSP50 mRNA 3'UTR driven luciferase activity reporting system,namely miR-937-5p?miR-2052?miR-1264 and miR-4709-3p.Then we transfected relative miRNAs mimics into A2780 cells which high-expression of TSP50,and investigated the effect of miRNAs on the expression of TSP50 in mRNA and protein levels by real-time PCR and western blot analysis,the results showed that four miRNAs can inhibit the expression of TSP50 in both mRNA and protein levels.Previous study revealed that TSP50 can promote cell proliferation,motility and invation in tumor cells.In order to detect the effect of miRNAs on the function of TSP50 in tumor development,we transfected four kinds of mi RNA mimics into high-expressing TSP50 cells A2780 and low-expressing TSP50 cells Hela,and analyzed the effects of miRNAs on cell proliferation by MTT and BrdU incorporation experiment,the results showed that miR-937-5p,miR-2052,miR-1264 and miR-4709-3p inhibited the proliferation of A2780 cells,but had no significant effect on Hela cells.Then we investigated the effects of miRNAs on cell motility and invation by wound healing and transwell assay,the results showed that four mi RNAs reduced the motility and invation of A2780 cells,however,Hela cells were not be significantly affected.Taken together,we screened and obtained four kinds of miRNAs targeting TSP50 mRNA 3'UTR,and detected the effects of miRNAs on the proliferation,mobility and invation of TSP50 high-expressing tumor cells.This study laid a foundation for further investigating the function of miRNAs on the effect of TSP50 in tumorigenesis and development,and also provided a novel approach for cancer treatment targeting TSP50.